The present disclosure relates to the field of biotechnology, and more specifically, to the capture and purification of recombinant proteins. BACKGROUND

Current techniques and tools for the production of recombinant proteins are often labor-intensive, time-consuming, costly, and error-prone. With each step requiring user manipulation, protein recovery, among other things, can suffer. SUMMARY

The present disclosure is based, at least in part, on the generation of novel filtration and chromatography pods for capturing and purifying recombinant proteins from a liquid cell culture. As elaborated on below and exemplified in the working examples, the filtration and chromatography pods offer several benefits over previous methods of purifying recombinant proteins, including, e.g., speed, ease of use, enhanced protein stability, eliminating the need for multiple culture vessels and other consumables, which allows for rapid capture and purification of the recombinant protein of interest. Other advantages of the presently claimed invention are described herein.

Accordingly, provided herein are filtration and chromatography pods that include a container being formed by a wall and having a top end and a bottom end, where the wall forms a seal with the bottom end, and one or more of the wall, the top end, and the bottom end include at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod. In some embodiments of any of the pods described herein, the bottom end includes at least one area covered by the semi-porous or porous material. In some embodiments of any of the pods described herein, the semi-porous or porous material is characterized by one or both of: (a) not allowing a cell to flow from the exterior of the pod into the interior of the pod and (b) retaining or substantially retaining the resin within the interior of the pod. In some embodiments, the semi- porous or porous material does not allow a cell to flow from the exterior of the pod into the interior of the pod. In some embodiments, the semi-porous or porous material retaining or substantially retaining the resin within the interior of the pod. In some embodiments of any of the pods described herein, the semi-porous or porous material is characterized by not allowing a cell to flow from the exterior of the pod into the interior of the pod and retaining or substantially retaining the resin within the interior of the pod. In some embodiments of any of the pods described herein, the cell is a mammalian cell. In some embodiments of any of the pods described herein, the cell is an insect cell or a yeast cell. In some embodiments of any of the pods described herein, the cell is a bacterial cell (e.g., an E. coli cell). In some embodiments of any of the pods, methods, kits, or any other aspects described herein, the cell is not a bacterial cell. In some embodiments, the semi-porous or porous material retains or substantially retains the resin within the interior of pod structure.

In some embodiments of any of the pods described herein, the material is a semi-permeable or porous membrane. In some embodiments of any of the pods described herein, the material is a semi-permeable or porous mesh. In some embodiments, the mesh is coated with a hydrophilic material or a hydrophobic material. In some embodiments of any of the pods described herein, the mesh is made of nylon or polyester. In some embodiments of any of the pods described herein, the material has a pore size of about 50 nm to about 100 µm. In some embodiments of any of the pods described herein, the material has a pore size of about 50 nm to about 50 µm. In some embodiments of any of the pods described herein, the material has a pore size of about 100 nm to about 50 μm. In some embodiments of any of the pods described herein, the material has a pore size of about 10 μm to about 50 μm. In some embodiments of any of the pods described herein, the material has a pore size of about 20 μm to about 50 μm. In some embodiments of any of the pods described herein, the material has a pore size of about 100 nm to about 30 µm. In some embodiments of any of the pods described herein, the material has a pore size of about 500 nm to about 20 µm. In some embodiments of any of the pods described herein, the material has a pore size of about 1 µm to about 10 µm. In some embodiments of any of the pods described herein, the material has a pore size of about 2 µm to about 5 µm.

In some embodiments of any of the pods described herein, the container has an interior volume of about 0.1 mL to about 500 mL. In some embodiments of any of the pods described herein, the container has an interior volume of about 0.1 mL to about 250 mL. In some embodiments of any of the pods described herein, the container has an interior volume of about 0.1 mL to about 100 mL. In some embodiments of any of the pods described herein, the bottom end includes at least one area covered by the material. In some embodiments of any of the pods described herein, the wall includes at least one area covered by the material. In some embodiments of any of the pods described herein, the bottom end and the wall each include at least one area covered by the material. In some embodiments of any of the pods described herein, the top end includes at least one area covered by the material. In some embodiments of any of the pods described herein, the top end and the bottom end each include at least one area covered by the material. In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² to about 25 cm ² . In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² to about 20 cm ² . In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² to about 10 cm ² . In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² to about 5 cm ² . In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² and 2.5 cm ² . In some embodiments of any of the pods described herein, each of the at least one area has a surface area of about 1 mm ² and about 1 cm ² .

In some embodiments of any of the pods described herein, the bottom end is flat. In some embodiments, the bottom end has a hemispherical shape. In some embodiments of any of the pods described herein, the bottom end has a conical shape. In some embodiments of any of the pods described herein, the wall has a cylindrical shape. In some embodiments of any of the pods described herein, the wall has a conical shape. In some embodiments of any of the pods described herein, the wall has a spherical shape. In some embodiments of any of the pods described herein, the wall has a hemispherical shape. In some embodiments of any of the pods described herein, the wall has an hourglass shape. In some embodiments of any of the pods described herein, the pod further includes a chromatography resin disposed in the interior of the container. In some embodiments of any of the pods described herein, the chromatography resin is a commercial chromatography resin. In some embodiments of any of the pods described herein, the chromatography resin is an affinity chromatography resin. In some embodiments of any of the pods described herein, the affinity chromatography resin is a protein A resin, a protein G resin, a protein L resin, a nickel resin, or a magnetic bead resin. In some embodiments of any of the pods described herein, the chromatography resin has a volume of about 0.1 mL to about 500 mL. In some embodiments of any of the pods described herein, the chromatography resin has a volume of about 0.1 mL to about 250 mL. In some embodiments of any of the pods described herein, the chromatography resin has a volume of about 0.1 mL to about 100 mL. In some embodiments of any of the pods described herein, the

chromatography resin has a volume of about 0.1 mL to about 50 mL. In some embodiments of any of the pods described herein, the chromatography resin has a sterility assurance level of about 1 x 10 ^-6 to about 1 x 10 ^-10 .

Also provided here are kits that include any of the pods described herein. Some embodiments of these kits can further include instructions for using any of the pods described herein. Some embodiments of any of the kits provided herein can further include a means for stably positioning the pod in the interior of a culture vessel. In some embodiments of any of the kits described herein, the culture vessel is shake flask, a shake tube, a bioreactor, or a multi-well plate. In some embodiments of any of the kits described herein, the bioreactor is a perfusion bioreactor. In some embodiments of any of the kits described herein, the bioreactor is a feed batch bioreactor.

Also provided herein are kits that include: (a) any of the pods described herein; and (b) one or more panels that can be secured over each of the one or more areas in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the wall is prevented.

Also provided herein are kits that include: (a) any of the pods described herein; and; (b) an outer sleeve that can be secured over the pod in order to cover the at least one area in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented. In some embodiments, the sleeve is an empty chromatography column. Also provided herein are kits that include: (a) a pod of any of the pods described herein; and (b) one or both of: an elution outlet that can be stably fixed onto the bottom end; and a buffer reservoir that can be stably fixed onto the top end. In some embodiments of any of the kits described herein, the kit includes the pod and the elution outlet. In some embodiments of any of the kits described herein, the kit includes the pod and the buffer reservoir. In some embodiments of any of the kits described herein, the kit includes the pod, the elution outlet, and the buffer reservoir. In some embodiments of any of the kits provided herein, the elution outlet is a funnel. In some embodiments of any of the kits provided herein, the elution outlet includes a flat surface having a port. In some embodiments of any of the kits provided herein, the elution outlet is configured to allow for regulation of the flow of a fluid through the elution outlet. In some embodiments of any of the kits provided herein, the elution outlet is not configured to allow for regulation of the flow of a fluid through the elution outlet. In some embodiments of any of the kits provided herein, the elution outlet can be screwed or clicked-on to the pod. In some embodiments of any of the kits provided herein, the elution outlet can be sealed to the pod using one or more (e.g., two or three) O-rings (e.g., fixed to the pod or the elution outlet).

Also provided herein are kits that include: (a) any of the pods described herein; and (b) a movable outer sleeve connected to the wall, such that rotation of the movable outer sleeve can cover the at least one area in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented. In some embodiments of any of the kits provided herein, the kit further includes a means for stably positioning the pod in the interior of a culture vessel.

Provided herein are kits that include: (a) any of the pods described herein; and (b) a means for stably positioning the long axis of the pod in a horizontal position in the interior of a culture vessel. Non-limiting examples of means for stably positioning the the pod in an interior of a culture vessel are shown in Figures 11-19, 36, and 37.

Also provided herein are kits that include (a) any of the pods described herein; (b) an inlet system having a top end, an elongated cylindrical body including a fluid conduit, and a bottom end, where the top end of the inlet system includes a fluid inlet and is configured for stable attachment to the top end of the pod and, when stably connected to the pod, allows for flow of a fluid into the pod through the fluid inlet and the fluid conduit, and out of the bottom end of the inlet system, and the elongated cylindrical body of the inlet system is configured to fit inside the wall of the pod; an (c) an outlet system including a fluid outlet that is configured for stable attachment to the bottom end of the pod, and when stably connected to the bottom end of the pod, allows for flow of a fluid out of the pod through the fluid outlet. In some embodiments of these kits, the top end of the inlet system and the top end of the pod include threading that allows for the stable attachment of the top end of the pod to the top end of the inlet system. In some embodiments of these kits, the threading in the top end of the inlet system or the threading in the top end of the pod comprises an O- ring. In some embodiments of these kits, the pod contains a chromatography resin (e.g., any of the exemplary types of chromatography resin described herein); and the inlet system and the pod are configured such that, when the inlet system and the pod are stably attached, the bottom end of the inlet system is proximal to the chromatography resin. In some embodiments of these kits, the interior surface of the pod and the bottom end of the inlet system are configured to allow for stable attachment of the pod and the bottom end of the inlet system. In some embodiments of these kits, the interior surface of the pod and the bottom end of the inlet system include threading that allows for stable attachment of the pod and the bottom end of the inlet system. In some embodiments of these kits, the threading in the interior surface of the pod or the threading in the bottom end of the inlet system includes an O-ring. In some embodiments of these kits, the outlet system and the bottom end of the pod include threading that allows for stable attachment of the outlet system to the bottom end of the pod. In some embodiments of these kits, the threading in the outlet system or the threading in the bottom end of the pod includes an O-ring. In some embodiments of these kits, the fluid inlet is capable of attachment to tubing. In some embodiments of these kits, the fluid outlet is capable of attachment to tubing. Some embodiments of these kits further include: (d) a means for stably positioning the long axis of the pod in a horizontal position in the interior of a culture vessel (e.g., any of the culture vessels described herein). Some embodiments of these kits further include: (d) a mean for stably positioning the long axis of the pod in a vertical position in the interior of a culture vessel (e.g., any of the culture vessels described herein).

Also provided herein are methods of capturing a recombinant protein produced by a liquid cell culture that include contacting any of the pods of described herein, with a liquid cell culture for a period of time, where the contacting step results in at least a portion of the at least one area contacting the liquid cell culture, and the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin. In some embodiments of any of the methods described herein, the liquid cell culture is disposed in a culture vessel.

Also provided herein are methods of purifying a recombinant protein produced by a liquid cell culture that include: (a) contacting any of the pods of described herein, with a liquid cell culture for a period of time, where the contacting step results in at least a portion of the at least one area contacting the liquid cell culture, and the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin; and (b) eluting the recombinant protein from the chromatography resin after the period of time, thereby purifying the recombinant protein.

In some embodiments of any of the methods provided herein, the method further includes, after (b), (c) performing one or more additional unit operations on the recombinant protein. In some embodiments of any of the methods provided herein, the method further includes, after (c), formulating the recombinant protein into a recombinant protein drug product. In some embodiments of any of the methods provided herein, the method further includes, washing the chromatography resin between step (a) and step (b). In some embodiments of any of the methods described herein, the contacting is performed under sterile conditions.

In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 2 weeks. In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 168 hours. In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 96 hours. In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 48 hours. In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 24 hours. In some embodiments of any of the methods described herein, the period of time is about 5 minutes to about 12 hours. In some embodiments of any of the methods described herein, the period of time occurs when the liquid cell culture is in its exponential growth phase. In some embodiments of any of the methods described herein, the period of time occurs when the liquid cell culture is in its stationary growth phase. In some embodiments, the contacting can occur when a liquid cell culture is in its death phase. In some embodiments of any of the methods described herein, at least 30% of the total surface area of the at least one area is in contact with the liquid cell culture over the period of time in step (a). In some embodiments of any of the methods described herein, at least 50% of the total surface area of the at least one area is in contact with the liquid cell culture over the period of time in step (a). In some embodiments of any of the methods described herein of any of the methods described herein, at least 75% of the total surface area of the at least one area is in contact with the liquid cell culture over the period of time in step (a). In some embodiments of any of the methods described herein, at least 90% of the total surface area of the at least one area is in contact with the liquid cell culture over the period of time in step (a).

In some embodiments of any of the methods described herein, the liquid cell culture is disposed in a culture vessel. In some embodiments of any of the methods described herein, the culture vessel is a plate (e.g., a multi-well plate), a shake tube, a shake flask, or a bioreactor. In some embodiments of any of the methods described herein, the bioreactor is a perfusion bioreactor. In some embodiments of any of the methods described herein, the bioreactor is a fed batch bioreactor. In some embodiments of any of the methods described herein, the liquid cell culture includes a bacterial cell, a yeast cell, insect cell, a plant cell, or a mammalian cell. In some embodiments of any of the methods described herein, the mammalian cell is a human cell. In some embodiments, the mammalian cell is a Chinese hamster ovary (CHO) cell or an NS0 cell.

In some embodiments of any of the methods described herein, the one or more additional unit operations is/are selected from the group of: viral inactivation, viral filtration, polishing, size-exclusion chromatography, cation exchange

chromatography, anion exchange chromatography, hydrophobic interaction chromatography, mixed mode chromatography, and ultrafiltration/diafiltration.

In some embodiments of any of the methods described herein, the recombinant protein is a secreted protein. In some embodiments of any of the methods described herein, the recombinant protein is any protein with or without a tag (e.g., a fluorescent tag, a radioisotope, a polyhistidine tag (HIS tag), a c-myc tag, a FLAG tag, a maltose- binding protein (MBP) tag, an albumin-binding protein (ABP) tag, a bacteriophage T7 epitope (T7-tag), a biotin-carboxy carrier protein (BCCP) tag, a bluetongue virus tag (B-tag), a calmodulin binding peptide tag, a cellulose binding domain tag, a chitin binding domain tag, a choline binding domain tag, a galactose-binding protein tag, a Glu-Glu epitope tag, a human influenza hemagglutinin tag, a Halo Tag®, a histidine affinity tag (HAT), a polyarginine tag (Asp-tag), a polyphenylalanine tag (Phe-tag), an S-TAG, a streptavidin-binding peptide, a streptavidin tag, a Strep-tag, an enzyme (e.g., alkaline phosphatase, chloramphenicol acetyl transferase (CAT), glutathione S- transferase), or a Fc tag. In some embodiments of any of the methods described herein, the recombinant protein is an antibody, an antigen-binding antibody fragment, an enzyme, or a fusion protein.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims. BRIEF DESCRIPTION OF DRAWINGS FIGs.1A to 1E show different exemplary pods and kits provided herein. FIG.2 shows an exemplary embodiment of a pod provided herein, where the longitudinal axis of the pod is approximately horizontal when the pod is disposed in a bioreactor.

FIG.3 shows the exemplary pod in FIG.2, after it has been removed from a bioreactor and rotated such that the longitudinal axis is approximately vertical and its bottom end connected to an elution outlet. Optionally, the top end of the pod can be connected to a buffer reservoir (not shown).

FIG.4 is an exemplary depiction of a pod as described herein disposed in a vessel (e.g., a bioreactor) with a rod transiting through holes in the pod and in the side walls of the vessel.

FIG.5 is a representative image of a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis comparing the traditional batch capture method (Trad.) and the Capture & Purify method (Cap. & Pure) under reduced and non-reduced conditions. E = 1μg of elution sample; FT = flow through sample; W = wash sample;

M = Biorad Precision Plus ladder marker;“-” = blank.

FIG.6 is a representative diagram comparing the experimental time line of the ÄKTA purification method, the traditional batch capture method and the Capture & Purify method, starting at day 4 post-transfection.

FIG.7 is a representative graph showing the high-performance liquid chromatography (HPLC) trace under four different conditions as outlined in Table 2.

FIG.8 is a representative graph showing the HPLC trace under four different conditions as outlined in Table 3.

FIG.9 is a representative graph showing the HPLC trace under two different conditions as outlined in Table 4.

FIGs.10-13 are two dimensional drawings of different exemplary means for stably positioning a pod in the interior of a culture vessel. The symmetrical two- pronged claw can hold any of the exemplary pods described herein in a culture vessel.

FIGs.14 and 15 are two dimensional drawing of a part of an exemplary means for stably positioning a pod in the interior of a culture vessel; the center of the part is able to attach to a symmetrical two-pronged claw that can hold any of the exemplary pods described herein in a culture vessel (as depicted in Figures 16-19).

FIGs.16-19 are two dimensional drawings of different exemplary means for stably positioning a pod in the interior of a culture vessel. The symmetrical two- pronged claw can hold any of the exemplary pods described herein in a culture vessel.

FIG.20 shows the overlaid elution profiles of 2 ^g injections of protein samples on an analytical TOSOH Supergel TSK mAb HTP column from concurrent purifications using a device similar to the one shown in Figures 27 and 28 or using traditional batch capture and elution. In both cases, clarified supernatant containing antibodies was exposed to protein A resin before washing and eluting from the protein A resin after removal of the device or free floating resin from the clarified supernatant.

FIG.21 shows the elution profile of a 2 ^g sample injected on an analytical TOSOH Supergel TSK mAb HTP column using the same clarified supernatant as Fig. 20 but using an AKTA system and proA resin contained within a packed column.

FIG.22 shows an exemplary pod provided herein.

FIG.23 shows an exemplary pod provided herein. FIGs.24-28 show exemplary kits provided herien.

FIGs.29-31 show exemplary pods provided herein.

FIGs.32 and 33 show elution profiles from a device depicted in Figures 27 and 28 containing a Protein A resin, which was exposed to an unfiltered supernatant and eluted using an ÄKTA system.

FIG.34 shows an exemplary adapter provided herein.

FIG.35 shows an exemplary adapter provided herein attached to an exemplary pod described herein.

FIG.36 and 37 show exemplary means for stably positioning the pod in the interior of a culture vessel. DETAILED DESCRIPTION

Provided herein are filtration and chromatography pods that include a container being formed by a wall and having a top end and a bottom end, where the wall forms a seal with the bottom end, and one or more of the wall, the top end, and the bottom end include at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod. Also provided herein are kits that include any of these pods, methods of using these pods to capture a recombinant protein produced in a liquid cell culture, and methods of using these pods to purify a recombinant protein produced by a liquid cell culture.

Non-limiting aspects of these pods, kits, and methods are described below, and can be used in any combination without limitation. Additional aspects of these pods, kits, and methods are known in the art.

The pods, kits, and methods provided herein provide for an unexpected decrease in the amount of time necessary to capture or purify a recombinant protein from a protein expression system (e.g., protein produced by a recombinant cell, such as a stable cell, expressing the recombinant protein). For example, a recombinant protein can be captured or purifying within about 5 minutes to about 20 hours, about 5 minutes to about 18 hours, about 5 minutes to about 16 hours, about 5 minutes to about 14 hours, about 5 minutes to about 12 hours, about 5 minutes to about 10 hours, about 5 minutes to about 8 hours, about 5 minutes to about 6 hours, about 5 minutes to about 4 hours, about 5 minutes to about 3 hours, about 5 minutes to about 2 hours, about 5 minutes to about 1 hour, about 5 minutes to about 30 minutes, about 5 minutes to about 20 minutes, about 5 minutes to about 10 minutes, about 10 minutes to about 20 hours, about 10 minutes to about 18 hours, about 10 minutes to about 16 hours, about 10 minutes to about 14 hours, about 10 minutes to about 12 hours, about 10 minutes to about 10 hours, about 10 minutes to about 8 hours, about 10 minutes to about 6 hours, about 10 minutes to about 4 hours, about 10 minutes to about 3 hours, about 10 minutes to about 2 hours, about 10 minutes to about 1 hour, about 10 minutes to about 30 minutes, about 10 minutes to about 20 minutes, about 20 minutes to about 20 hours, about 20 minutes to about 18 hours, about 20 minutes to about 16 hours, about 20 minutes to about 14 hours, about 20 minutes to about 12 hours, about 20 minutes to about 10 hours, about 20 minutes to about 8 hours, about 20 minutes to about 6 hours, about 20 minutes to about 4 hours, about 20 minutes to about 3 hours, about 20 minutes to about 2 hours, about 20 minutes to about 1 hour, about 20 minutes to about 30 minutes, about 30 minutes to about 20 hours, about 30 minutes to about 18 hours, about 30 minutes to about 16 hours, about 30 minutes to about 14 hours, about 30 minutes to about 12 hours, about 30 minutes to about 10 hours, about 30 minutes to about 8 hours, about 30 minutes to about 6 hours, about 30 minutes to about 4 hours, about 30 minutes to about 3 hours, about 30 minutes to about 2 hours, about 30 minutes to about 1 hour, about 1 hour to about 20 hours, about 1 hour to about 18 hours, about 1 hour to about 16 hours, about 1 hour to about 14 hours, about 1 hour to about 12 hours, about 1 hour to about 10 hours, about 1 hour to about 8 hours, about 1 hour to about 6 hours, about 1 hour to about 4 hours, about 1 hour to about 3 hours, about 1 hour to about 2 hours, about 2 hours to about 20 hours, about 2 hours to about 18 hours, about 2 hours to about 16 hours, about 2 hours to about 14 hours, about 2 hours to about 12 hours, about 2 hours to about 10 hours, about 2 hours to about 8 hours, about 2 hours to about 6 hours, about 2 hours to about 4 hours, about 2 hours to about 3 hours, about 3 hours to about 20 hours, about 3 hours to about 18 hours, about 3 hours to about 16 hours, about 3 hours to about 14 hours, about 3 hours to about 12 hours, about 3 hours to about 10 hours, about 3 hours to about 8 hours, about 3 hours to about 6 hours, about 3 hours to about 4 hours, about 4 hours to about 20 hours, about 4 hours to about 18 hours, about 4 hours to about 16 hours, about 4 hours to about 14 hours, about 4 hours to about 12 hours, about 4 hours to about 10 hours, about 4 hours to about 8 hours, about 4 hours to about 6 hours, about 6 hours to about 20 hours, about 6 hours to about 18 hours, about 6 hours to about 16 hours, about 6 hours to about 14 hours, about 6 hours to about 12 hours, about 6 hours to about 10 hours, about 6 hours to about 8 hours, about 8 hours to about 20 hours, about 8 hours to about 18 hours, about 8 hours to about 16 hours, about 8 hours to about 14 hours, about 8 hours to about 12 hours, about 8 hours to about 10 hours, about 10 hours to about 20 hours, about 10 hours to about 18 hours, about 10 hours to about 16 hours, about 10 hours to about 14 hours, about 10 hours to about 12 hours, about 12 hours to about 20 hours, about 12 hours to about 18 hours, about 12 hours to about 16 hours, about 12 hours to about 14 hours, about 14 hours to about 20 hours, about 14 hours to about 18 hours, about 14 hours to about 16 hours, about 16 hours to about 20 hours, about 16 hours to about 18 hours, or about 18 hours to about 20 hours. Pods

Provided herein are filtration and chromatography pods that include a container being formed by a wall and having a top end and a bottom end, where the wall forms a seal with the bottom end, and one or more of the wall, the top end, and the bottom end include at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod. The semi-porous or porous material can be attached to the wall, top end, and/or bottom end of the pod by heating sealing. Other means for attaching the semi- porous or porous material to the wall, top end, and/or bottom end of the pod are known in the art. Non-limiting examples of the pods and kits provided herein are shown in Figures 1A to 1E, Figure 2, Figure 3, Figure 4, Figures 22-31, and Figure 35. In some embodiments, pod designs, such as those set forth in Figure 1A, may optionally include one or more panels adapted to cover some or all of the surface of the sides of the pod which are formed by the semi-porous or porous material.

Attachment of such covers can further facilitate subsequent purification steps. Semi-Porous / Porous Materials

In some embodiments of any of the pods described herein, the semi-porous or porous material retains or substantially retains the resin within the pod structure. In some examples of any of the pods described herein, the semi-porous or porous material is characterized by one or both of: (a) not allowing a cell to flow from the exterior of the pod into the interior of the pod and (b) retaining or substantially retaining the resin within the interior of the pod (c) is coated with a hydrophilic or hydrophobic coating. In some embodiments, the semi-porous or porous material does not allow a cell to flow from the exterior of the pod into the interior of the pod. In some embodiments, the semi-porous or porous material retaining or substantially retaining the resin within the interior of the pod. In some embodiments of any of the pods described herein, the semi-porous or porous material is characterized by not allowing a cell to flow from the exterior of the pod into the interior of the pod and retaining or substantially retaining the resin within the interior of the pod.

In some examples of any of the pods described herein, the semi-porous or porous material is characterized by one or both of: (a) not allowing a mammalian cell to flow from the exterior of the pod into the interior of the pod and (b) retaining or substantially retaining the resin within the interior of the pod. In some examples of any of the pods described herein, the semi-porous or porous material does not allow a mammalian cell to reversibly flow from the exterior of the pod into the interior of the pod. In some examples of any of the pods described herein, the semi-porous or porous material is characterized by one or both of: (a) not allowing an insect cell to flow from the exterior of the pod into the interior of the pod and (b) retaining or substantially retaining the resin within the interior of the pod. In some examples of any of the pods described herein, the semi-porous or porous material does not allow an insect cell or a yeast cell to reversibly flow from the exterior of the pod into the interior of the pod. In some examples of any of the pods described herein, the semi- porous or porous material does not allow a bacterial cell to reversibly flow from the exterior of the pod into the interior of the pod.

In some examples, the material is a semi-permeable or porous membrane. In some examples, the material is a semi-permeable or porous mesh (e.g., a nylon mesh).

In some examples, the material can be include (at least in part) one or more of the following: polyester, polyester with a hydrophilic coating, cellulose acetate, cellulose nitrate, polyethersulfone (PES), polystyrene, polypropylene, acrylic copolymer, polyvinyl chloride (PVC), polyvinylidene difluoride (PVDF), polytetrafluoroethylene (PTFE), and polycarbonate. In some embodiments, the material is a precision woven polymer (e.g., any precision woven polymer). In some embodiments, the material is a Sefar film (see, e.g., Sefar’s U.S. website, www.sefar.us). In some embodiments, the material is Petex mesh sold by Sefar.

In some embodiments of any of the pods described herein, the material has a pore size or average pore size of about 50 nm to about 50 µm, about 50 nm to about 45 µm, about 50 nm to about 40 µm, about 50 nm to about 35 µm, about 50 nm to about 30 µm, about 50 nm to about 25 µm, about 50 nm to about 20 µm, about 50 nm to about 18 µm, about 50 nm to about 16 µm, about 50 nm to about 14 µm, about 50 nm to about 12 µm, about 50 nm to about 10 µm, about 50 nm to about 8 µm, about 50 nm to about 6 µm, about 50 nm to about 5 µm, about 50 nm to about 4 µm, about 50 nm to about 3 µm, about 50 nm to about 2 µm, about 50 nm to about 1 µm, about 50 nm to about 500 nm, about 50 nm to about 100 nm, about 100 nm to about 50 µm, about 100 nm to about 45 µm, about 100 nm to about 40 µm, about 100 nm to about 35 µm, about 100 nm to about 30 µm, about 100 nm to about 25 µm, about 100 nm to about 20 µm, about 100 nm to about 18 µm, about 100 nm to about 16 µm, about 100 nm to about 14 µm, about 100 nm to about 12 µm, about 100 nm to about 10 µm, about 100 nm to about 8 µm, about 100 nm to about 6 µm, about 100 nm to about 5 µm, about 100 nm to about 4 µm, about 100 nm to about 3 µm, about 100 nm to about 2 µm, about 100 nm to about 1 µm, about 100 nm to about 500 nm, about 500 nm to about 50 µm, about 500 nm to about 45 µm, about 500 nm to about 40 µm, about 500 nm to about 35 µm, about 500 nm to about 30 µm, about 500 nm to about 25 µm, about 500 nm to about 20 µm, about 500 nm to about 18 µm, about 500 nm to about 16 µm, about 500 nm to about 14 µm, about 500 nm to about 12 µm, about 500 nm to about 10 µm, about 500 nm to about 8 µm, about 500 nm to about 6 µm, about 500 nm to about 5 µm, about 500 nm to about 4 µm, about 500 nm to about 3 µm, about 500 nm to about 2 µm, about 500 nm to about 1 µm, about 1 µm to about 50 µm, about 1 µm to about 45 µm, about 1 µm to about 40 µm, about 1 µm to about 35 µm, about 1 µm to about 30 µm, about 1 µm to about 25 µm, about 1 µm to about 20 µm, about 1 µm to about 18 µm, about 1 µm to about 16 µm, about 1 µm to about 14 µm, about 1 µm to about 12 µm, about 1 µm to about 10 µm, about 1 µm to about 8 µm, about 1 µm to about 6 µm, about 1 µm to about 5 µm, about 1 µm to about 4 µm, about 1 µm to about 3 µm, about 1 µm to about 2 µm, about 2 µm to about 50 µm, about 2 µm to about 45 µm, about 2 µm to about 40 µm, about 2 µm to about 35 µm, about 2 µm to about 30 µm, about 2 µm to about 25 µm, about 2 µm to about 20 µm, about 2 µm to about 18 µm, about 2 µm to about 16 µm, about 2 µm to about 14 µm, about 2 µm to about 12 µm, about 2 µm to about 10 µm, about 2 µm to about 8 µm, about 2 µm to about 6 µm, about 2 µm to about 5 µm, about 2 µm to about 4 µm, about 2 µm to about 3 µm, about 3 µm to about 50 µm, about 3 µm to about 45 µm, about 3 µm to about 40 µm, about 3 µm to about 35 µm, about 3 µm to about 30 µm, about 3 µm to about 25 µm, about 3 µm to about 20 µm, about 3 µm to about 18 µm, about 3 µm to about 16 µm, about 3 µm to about 14 µm, about 3 µm to about 12 µm, about 3 µm to about 10 µm, about 3 µm to about 8 µm, about 3 µm to about 6 µm, about 3 µm to about 5 µm, about 3 µm to about 4 µm, about 4 µm to about 50 µm, about 4 µm to about 45 µm, about 4 µm to about 40 µm, about 4 µm to about 35 µm, about 4 µm to about 30 µm, about 4 µm to about 25 µm, about 4 µm to about 20 µm, about 4 µm to about 18 µm, about 4 µm to about 16 µm, about 4 µm to about 14 µm, about 4 µm to about 12 µm, about 4 µm to about 10 µm, about 4 µm to about 8 µm, about 4 µm to about 6 µm, about 4 µm to about 5 µm, about 5 µm to about 50 µm, about 5 µm to about 45 µm, about 5 µm to about 40 µm, about 5 µm to about 35 µm, about 5 µm to about 30 µm, about 5 µm to about 25 µm, about 5 µm to about 20 µm, about 5 µm to about 18 µm, about 5 µm to about 16 µm, about 5 µm to about 14 µm, about 5 µm to about 12 µm, about 5 µm to about 10 µm, about 5 µm to about 8 µm, about 5 µm to about 6 µm, about 6 µm to about 50 µm, about 6 µm to about 45 µm, about 6 µm to about 40 µm, about 6 µm to about 35 µm, about 6 µm to about 30 µm, about 6 µm to about 25 µm, about 6 µm to about 20 µm, about 6 µm to about 18 µm, about 6 µm to about 16 µm, about 6 µm to about 14 µm, about 6 µm to about 12 µm, about 6 µm to about 10 µm, about 6 µm to about 8 µm, about 8 µm to about 50 µm, about 8 µm to about 45 µm, about 8 µm to about 40 µm, about 8 µm to about 35 µm, about 8 µm to about 30 µm, about 8 µm to about 25 µm, about 8 µm to about 20 µm, about 8 µm to about 18 µm, about 8 µm to about 16 µm, about 8 µm to about 14 µm, about 8 µm to about 12 µm, about 8 µm to about 10 µm, about 10 µm to about 50 µm, about 10 µm to about 45 µm, about 10 µm to about 40 µm, about 10 µm to about 35 µm, about 10 µm to about 30 µm, about 10 µm to about 25 µm, about 10 µm to about 20 µm, about 10 µm to about 18 µm, about 10 µm to about 16 µm, about 10 µm to about 14 µm, about 10 µm to about 12 µm, about 12 µm to about 50 µm, about 12 µm to about 45 µm, about 12 µm to about 40 µm, about 12 µm to about 35 µm, about 12 µm to about 30 µm, about 12 µm to about 25 µm, about 12 µm to about 20 µm, about 12 µm to about 18 µm, about 12 µm to about 16 µm, about 12 µm to about 14 µm, about 14 µm to about 50 µm, about 14 µm to about 45 µm, about 14 µm to about 40 µm, about 14 µm to about 35 µm, about 14 µm to about 30 µm, about 14 µm to about 25 µm, about 14 µm to about 20 µm, about 14 µm to about 18 µm, about 14 µm to about 16 µm, about 16 µm to about 50 µm, about 16 µm to about 45 µm, about 16 µm to about 40 µm, about 16 µm to about 35 µm, about 16 µm to about 30 µm, about 16 µm to about 25 µm, about 16 µm to about 20 µm, about 16 µm to about 18 µm, about 18 µm to about 50 µm, about 18 µm to about 45 µm, about 18 µm to about 40 µm, about 18 µm to about 35 µm, about 18 µm to about 30 µm, about 18 µm to about 25 µm, about 18 µm to about 20 µm, about 20 µm to about 50 µm, about 20 µm to about 45 µm, about 20 µm to about 40 µm, about 20 µm to about 35 µm, about 20 µm to about 30 µm, about 20 µm to about 25 µm, about 25 µm to about 50 µm, about 25 µm to about 45 µm, about 25 µm to about 40 µm, about 25 µm to about 35 µm, about 25 µm to about 30 µm, about 30 µm to about 50 µm, about 30 µm to about 45 µm, about 30 µm to about 40 µm, about 30 µm to about 35 µm, about 35 µm to about 50 µm, about 35 µm to about 45 µm, about 35 µm to about 40 µm, about 40 µm to about 50 µm, about 40 µm to about 45 µm, or about 45 µm to about 50 µm. In some embodiments of any of the pods described herein, the material has a pore size or average pore size of about 30 μm to about 70 μm, about 30 μm to about 68 μm, about 30 μm to about 66 μm, about 30 μm to about 64 μm, about 30 μm to about 62 μm, about 30 μm to about 60 μm, about 30 μm to about 58 μm, about 30 μm to about 56 μm, about 30 μm to about 54 μm, about 30 μm to about 52 μm, about 30 μm to about 50 μm, about 30 μm to about 48 μm, about 30 μm to about 46 μm, about 30 μm to about 44 μm, about 30 μm to about 42 μm, about 30 μm to about 40 μm, about 30 μm to about 38 μm, about 30 μm to about 36 μm, about 30 μm to about 34 μm, about 30 μm to about 32 μm, about 32 μm to about 70 μm, about 32 μm to about 68 μm, about 32 μm to about 66 μm, about 32 μm to about 64 μm, about 32 μm to about 62 μm, about 32 μm to about 60 μm, about 32 μm to about 58 μm, about 32 μm to about 56 μm, about 32 μm to about 54 μm, about 32 μm to about 52 μm, about 32 μm to about 50 μm, about 32 μm to about 48 μm, about 32 μm to about 46 μm, about 32 μm to about 44 μm, about 32 μm to about 42 μm, about 32 μm to about 40 μm, about 32 μm to about 38 μm, about 32 μm to about 36 μm, about 32 μm to about 34 μm, about 34 μm to about 70 μm, about 34 μm to about 68 μm, about 34 μm to about 66 μm, about 34 μm to about 64 μm, about 34 μm to about 62 μm, about 34 μm to about 60 μm, about 34 μm to about 58 μm, about 34 μm to about 56 μm, about 34 μm to about 54 μm, about 34 μm to about 52 μm, about 34 μm to about 50 μm, about 34 μm to about 48 μm, about 34 μm to about 46 μm, about 34 μm to about 44 μm, about 34 μm to about 42 μm, about 34 μm to about 40 μm, about 34 μm to about 38 μm, about 34 μm to about 36 μm, about 36 μm to about 70 μm, about 36 μm to about 68 μm, about 36 μm to about 66 μm, about 36 μm to about 64 μm, about 36 μm to about 62 μm, about 36 μm to about 60 μm, about 36 μm to about 58 μm, about 36 μm to about 56 μm, about 36 μm to about 54 μm, about 36 μm to about 52 μm, about 36 μm to about 50 μm, about 36 μm to about 48 μm, about 36 μm to about 46 μm, about 36 μm to about 44 μm, about 36 μm to about 42 μm, about 36 μm to about 40 μm, about 36 μm to about 38 μm, about 38 μm to about 70 μm, about 38 μm to about 68 μm, about 38 μm to about 66 μm, about 38 μm to about 64 μm, about 38 μm to about 62 μm, about 38 μm to about 60 μm, about 38 μm to about 58 μm, about 38 μm to about 56 μm, about 38 μm to about 54 μm, about 38 μm to about 52 μm, about 38 μm to about 50 μm, about 38 μm to about 48 μm, about 38 μm to about 46 μm, about 38 μm to about 44 μm, about 38 μm to about 42 μm, about 38 μm to about 40 μm, about 40 μm to about 70 μm, about 40 μm to about 68 μm, about 40 μm to about 66 μm, about 40 μm to about 64 μm, about 40 μm to about 62 μm, about 40 μm to about 60 μm, about 40 μm to about 58 μm, about 40 μm to about 56 μm, about 40 μm to about 54 μm, about 40 μm to about 52 μm, about 40 μm to about 50 μm, about 40 μm to about 48 μm, about 40 μm to about 46 μm, about 40 μm to about 44 μm, about 40 μm to about 42 μm, about 42 μm to about 70 μm, about 42 μm to about 68 μm, about 42 μm to about 66 μm, about 42 μm to about 64 μm, about 42 μm to about 62 μm, about 42 μm to about 60 μm, about 42 μm to about 58 μm, about 42 μm to about 56 μm, about 42 μm to about 54 μm, about 42 μm to about 52 μm, about 42 μm to about 50 μm, about 42 μm to about 48 μm, about 42 μm to about 46 μm, about 42 μm to about 44 μm, about 44 μm to about 70 μm, about 44 μm to about 68 μm, about 44 μm to about 66 μm, about 44 μm to about 64 μm, about 44 μm to about 62 μm, about 44 μm to about 60 μm, about 44 μm to about 58 μm, about 44 μm to about 56 μm, about 44 μm to about 54 μm, about 44 μm to about 52 μm, about 44 μm to about 50 μm, about 44 μm to about 48 μm, about 44 μm to about 46 μm, about 46 μm to about 70 μm, about 46 μm to about 68 μm, about 46 μm to about 66 μm, about 46 μm to about 64 μm, about 46 μm to about 62 μm, about 46 μm to about 60 μm, about 46 μm to about 58 μm, about 46 μm to about 56 μm, about 46 μm to about 54 μm, about 46 μm to about 52 μm, about 46 μm to about 50 μm, about 46 μm to about 48 μm, about 48 μm to about 70 μm, about 48 μm to about 68 μm, about 48 μm to about 66 μm, about 48 μm to about 64 μm, about 48 μm to about 62 μm, about 48 μm to about 60 μm, about 48 μm to about 58 μm, about 48 μm to about 56 μm, about 48 μm to about 54 μm, about 48 μm to about 52 μm, about 48 μm to about 50 μm, about 50 μm to about 70 μm, about 50 μm to about 68 μm, about 50 μm to about 66 μm, about 50 μm to about 64 μm, about 50 μm to about 62 μm, about 50 μm to about 60 μm, about 50 μm to about 58 μm, about 50 μm to about 56 μm, about 50 μm to about 54 μm, about 50 μm to about 52 μm, about 52 μm to about 70 μm, about 52 μm to about 68 μm, about 52 μm to about 66 μm, about 52 μm to about 64 μm, about 52 μm to about 62 μm, about 52 μm to about 60 μm, about 52 μm to about 58 μm, about 52 μm to about 56 μm, about 52 μm to about 54 μm, about 54 μm to about 70 μm, about 54 μm to about 68 μm, about 54 μm to about 66 μm, about 54 μm to about 64 μm, about 54 μm to about 62 μm, about 54 μm to about 60 μm, about 54 μm to about 58 μm, about 54 μm to about 56 μm, about 56 μm to about 70 μm, about 56 μm to about 68 μm, about 56 μm to about 66 μm, about 56 μm to about 64 μm, about 56 μm to about 62 μm, about 56 μm to about 60 μm, about 56 μm to about 58 μm, about 58 μm to about 70 μm, about 58 μm to about 68 μm, about 58 μm to about 66 μm, about 58 μm to about 64 μm, about 58 μm to about 62 μm, about 58 μm to about 60 μm, about 60 μm to about 70 μm, about 60 μm to about 68 μm, about 60 μm to about 66 μm, about 60 μm to about 64 μm, about 60 μm to about 62 μm, about 62 μm to about 70 μm, about 62 μm to about 68 μm, about 62 μm to about 66 μm, about 62 μm to about 64 μm, about 64 μm to about 70 μm, about 64 μm to about 68 μm, about 64 μm to about 66 μm, about 66 μm to about 70 μm, about 66 μm to about 68 μm, or about 68 μm to about 70 μm. Areas Covered by Semi-Porous/ Porous Materials

In some embodiments of the pods, one or more of the wall, the top end, and the bottom end include at least one (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod. In some embodiments of these pods, the pod includes: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 areas covered by a semi-porous or porous material (that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod) in the wall, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 areas covered by a semi-porous or porous material (that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod) in the bottom end, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 areas covered by a semi-porous or porous material (that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod) in the top end.

In some examples of the pods described herein, the wall includes at least one area covered by the material. In some examples of the pods described herein, the bottom end and the wall each include at least one area covered by the material. In some examples, the top end comprises at least one area covered by the material. In some examples, the top end and the bottom end each include at least one area covered by the material.

In some embodiments of the pods described herein, the total number of areas covered by a semi-porous or porous material (that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod) in one or more of the wall, the top end, and the bottom end is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30. In some embodiments of the pods described herein, the total number of areas covered by a semi-porous or porous material (that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod) in one or more of the wall, the top end, and the bottom end is 1 to 30, 1 to 28, 1 to 26, 1 to 24, 1 to 22, 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, 2 to 30, 2 to 28, 2 to 26, 2 to 24, 2 to 22, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 2 or 3, 3 to 30, 3 to 28, 3 to 26, 3 to 24, 3 to 22, 3 to 20, 3 to 18, 3 to 16, 3 to 14, 3 to 12, 3 to 10, 3 to 8, 3 to 6, 3 or 4, 4 to 30, 4 to 28, 4 to 26, 4 to 24, 4 to 22, 4 to 20, 4 to 18, 4 to 16, 4 to 14, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 6 to 30, 6 to 28, 6 to 26, 6 to 24, 6 to 22, 6 to 20, 6 to 18, 6 to 16, 6 to 14, 6 to 12, 6 to 10, 6 to 8, 8 to 30, 8 to 28, 8 to 26, 8 to 24, 8 to 22, 8 to 20, 8 to 18, 8 to 16, 8 to 14, 8 to 12, 8 to 10, 10 to 30, 10 to 28, 10 to 26, 10 to 24, 10 to 22, 10 to 20, 10 to 18, 10 to 16, 10 to 14, 10 to 12, 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 22, 12 to 20, 12 to 18, 12 to 16, 12 to 14, 14 to 30, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20, 14 to 18, 14 to 16, 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20, 16 to 18, 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, 18 to 20, 20 to 30, 20 to 28, 20 to 26, 20 to 24, 22 to 22, 22 to 30, 22 to 28, 22 to 26, 22 to 24, 24 to 30, 24 to 28, 24 to 26, 26 to 30, 26 to 28, or 28 to 30.

In some embodiments of any of the pods described herein, one or more of the at least one area covered by a semi-porous or porous material is circular, ellipsoidal, square, rectangular, elongated (e.g., an elongated line or rectangle), star-shaped, zig- zag shaped, or serpentine shaped. In some embodiments, one or more (e.g., each) of the at least one areas covered by the semi-porous or porous material has a surface area of about 0.5 mm ² to about 60 cm ² , about 0.5 mm ² to about 58 cm ² , about 0.5 mm ² to about 56 cm ² , about 0.5 mm ² to about 54 cm ² , about 0.5 mm ² to about 52 cm ² , about 0.5 mm ² to about 50 cm ² , about 0.5 mm ² to about 48 cm ² , about 0.5 mm ² to about 46 cm ² , about 0.5 mm ² to about 44 cm ² , about 0.5 mm ² to about 42 cm ² , about 0.5 mm ² to about 40 cm ² , about 0.5 mm ² to about 38 cm ² , about 0.5 mm ² to about 36 cm ² , about 0.5 mm ² to about 34 cm ² , about 0.5 mm ² to about 32 cm ² , 0.5 mm ² to about 30 cm ² , about 0.5 mm ² to about 28 cm ² , about 0.5 mm ² to about 26 cm ² , about 0.5 mm ² to about 24 cm ² , about 0.5 mm ² to about 22 cm ² , about 0.5 mm ² to about 20 cm ² , about 0.5 mm ² to about 18 cm ² , about 0.5 mm ² to about 16 cm ² , about 0.5 mm ² to about 14 cm ² , about 0.5 mm ² to about 12 cm ² , about 0.5 mm ² to about 10 cm ² , about 0.5 mm ² to about 8 cm ² , about 0.5 mm ² to about 6 cm ² , about 0.5 mm ² to about 5 cm ² , about 0.5 mm ² to about 4 cm ² , about 0.5 mm ² to about 3 cm ² , about 0.5 mm ² to about 2 cm ² , about 0.5 mm ² to about 1 cm ² , about 0.5 mm ² to about 5 mm ² , about 0.5 mm ² to about 2 mm ² , about 0.5 mm ² to about 1 mm ² , about 1 mm ² to about 60 cm ² , about 1 mm ² to about 58 cm ² , about 1 mm ² to about 56 cm ² , about 1 mm ² to about 54 cm ² , about 1 mm ² to about 52 cm ² , about 1 mm ² to about 50 cm ² , about 1 mm ² to about 48 cm ² , about 1 mm ² to about 46 cm ² , about 1 mm ² to about 44 cm ² , about 1 mm ² to about 42 cm ² , about 1 mm ² to about 40 cm ² , about 1 mm ² to about 38 cm ² , about 1 mm ² to about 36 cm ² , about 1 mm ² to about 34 cm ² , about 1 mm ² to about 32 cm ² , about 1 mm ² to about 30 cm ² , about 1 mm ² to about 28 cm ² , about 1 mm ² to about 26 cm ² , about 1 mm ² to about 24 cm ² , about 1 mm ² to about 22 cm ² , about 1 mm ² to about 20 cm ² , about 1 mm ² to about 18 cm ² , about 1 mm ² to about 16 cm ² , about 1 mm ² to about 14 cm ² , about 1 mm ² to about 12 cm ² , about 1 mm ² to about 10 cm ² , about 1 mm ² to about 8 cm ² , about 1 mm ² to about 6 cm ² , about 1 mm ² to about 5 cm ² , about 1 mm ² to about 4 cm ² , about 1 mm ² to about 3 cm ² , about 1 mm ² to about 2 cm ² , about 1 mm ² to about 1 cm ² , about 1 mm ² to about 5 mm ² , about 1 mm ² to about 2 mm ² , about 2 mm ² to about 60 cm ² , about 2 mm ² to about 58 cm ² , about 2 mm ² to about 56 cm ² , about 2 mm ² to about 54 cm ² , about 2 mm ² to about 52 cm ² , about 2 mm ² to about 50 cm ² , about 2 mm ² to about 48 cm ² , about 2 mm ² to about 46 cm ² , about 2 mm ² to about 44 cm ² , about 2 mm ² to about 42 cm ² , about 2 mm ² to about 40 cm ² , about 2 mm ² to about 38 cm ² , about 2 mm ² to about 36 cm ² , about 2 mm ² to about 34 cm ² , about 2 mm ² to about 32 cm ² , about 2 mm ² to about 30 cm ² , about 2 mm ² to about 28 cm ² , about 2 mm ² to about 26 cm ² , about 2 mm ² to about 24 cm ² , about 2 mm ² to about 22 cm ² , about 2 mm ² to about 20 cm ² , about 2 mm ² to about 18 cm ² , about 2 mm ² to about 16 cm ² , about 2 mm ² to about 14 cm ² , about 2 mm ² to about 12 cm ² , about 2 mm ² to about 10 cm ² , about 2 mm ² to about 8 cm ² , about 2 mm ² to about 6 cm ² , about 2 mm ² to about 5 cm ² , about 2 mm ² to about 4 cm ² , about 2 mm ² to about 3 cm ² , about 2 mm ² to about 2 cm ² , about 2 mm ² to about 1 cm ² , about 2 mm ² to about 5 mm ² , about 5 mm ² to about 60 cm ² , about 5 mm ² to about 58 cm ² , about 5 mm ² to about 56 cm ² , about 5 mm ² to about 54 cm ² , about 5 mm ² to about 52 cm ² , about 5 mm ² to about 50 cm ² , about 5 mm ² to about 48 cm ² , about 5 mm ² to about 46 cm ² , about 5 mm ² to about 44 cm ² , about 5 mm ² to about 42 cm ² , about 5 mm ² to about 40 cm ² , about 5 mm ² to about 38 cm ² , about 5 mm ² to about 36 cm ² , about 5 mm ² to about 34 cm ² , about 5 mm ² to about 32 cm ² , about 5 mm ² to about 30 cm ² , about 5 mm ² to about 28 cm ² , about 5 mm ² to about 26 cm ² , about 5 mm ² to about 24 cm ² , about 5 mm ² to about 22 cm ² , about 5 mm ² to about 20 cm ² , about 5 mm ² to about 18 cm ² , about 5 mm ² to about 16 cm ² , about 5 mm ² to about 14 cm ² , about 5 mm ² to about 12 cm ² , about 5 mm ² to about 10 cm ² , about 5 mm ² to about 8 cm ² , about 5 mm ² to about 6 cm ² , about 5 mm ² to about 5 cm ² , about 5 mm ² to about 4 cm ² , about 5 mm ² to about 3 cm ² , about 5 mm ² to about 2 cm ² , about 5 mm ² to about 1 cm ² , about 1 cm ² to about 60 cm ² , about 1 cm ² to about 58 cm ² , about 1 cm ² to about 56 cm ² , about 1 cm ² to about 54 cm ² , about 1 cm ² to about 52 cm ² , about 1 cm ² to about 50 cm ² , about 1 cm ² to about 48 cm ² , about 1 cm ² to about 46 cm ² , about 1 cm ² to about 44 cm ² , about 1 cm ² to about 42 cm ² , about 1 cm ² to about 40 cm ² , about 1 cm ² to about 38 cm ² , about 1 cm ² to about 36 cm ² , about 1 cm ² to about 34 cm ² , about 1 cm ² to about 32 cm ² , about 1 cm ² to about 30 cm ² , about 1 cm ² to about 28 cm ² , about 1 cm ² to about 26 cm ² , about 1 cm ² to about 24 cm ² , about 1 cm ² to about 22 cm ² , about 1 cm ² to about 20 cm ² , about 1 cm ² to about 18 cm ² , about 1 cm ² to about 16 cm ² , about 1 cm ² to about 14 cm ² , about 1 cm ² to about 12 cm ² , about 1 cm ² to about 10 cm ² , about 1 cm ² to about 8 cm ² , about 1 cm ² to about 6 cm ² , about 1 cm ² to about 5 cm ² , about 1 cm ² to about 4 cm ² , about 1 cm ² to about 3 cm ² , about 1 cm ² to about 2 cm ² , about 2 cm ² to about 60 cm ² , about 2 cm ² to about 58 cm ² , about 2 cm ² to about 56 cm ² , about 2 cm ² to about 54 cm ² , about 2 cm ² to about 52 cm ² , about 2 cm ² to about 50 cm ² , about 2 cm ² to about 48 cm ² , about 2 cm ² to about 46 cm ² , about 2 cm ² to about 44 cm ² , about 2 cm ² to about 42 cm ² , about 2 cm ² to about 40 cm ² , about 2 cm ² to about 38 cm ² , about 2 cm ² to about 36 cm ² , about 2 cm ² to about 34 cm ² , about 2 cm ² to about 32 cm ² , about 2 cm ² to about 30 cm ² , about 2 cm ² to about 28 cm ² , about 2 cm ² to about 26 cm ² , about 2 cm ² to about 24 cm ² , about 2 cm ² to about 22 cm ² , about 2 cm ² to about 20 cm ² , about 2 cm ² to about 18 cm ² , about 2 cm ² to about 16 cm ² , about 2 cm ² to about 14 cm ² , about 2 cm ² to about 12 cm ² , about 2 cm ² to about 10 cm ² , about 2 cm ² to about 8 cm ² , about 2 cm ² to about 6 cm ² , about 2 cm ² to about 5 cm ² , about 2 cm ² to about 4 cm ² , about 2 cm ² to about 3 cm ² ,about 3 cm ² to about 60 cm ² , about 3 cm ² to about 58 cm ² , about 3 cm ² to about 56 cm ² , about 3 cm ² to about 54 cm ² , about 3 cm ² to about 52 cm ² , about 3 cm ² to about 50 cm ² , about 3 cm ² to about 48 cm ² , about 3 cm ² to about 46 cm ² , about 3 cm ² to about 44 cm ² , about 3 cm ² to about 42 cm ² , about 3 cm ² to about 40 cm ² , about 3 cm ² to about 38 cm ² , about 3 cm ² to about 36 cm ² , about 3 cm ² to about 34 cm ² , about 3 cm ² to about 32 cm ² , about 3 cm ² to about 30 cm ² , about 3 cm ² to about 28 cm ² , about 3 cm ² to about 26 cm ² , about 3 cm ² to about 24 cm ² , about 3 cm ² to about 22 cm ² , about 3 cm ² to about 20 cm ² , about 3 cm ² to about 18 cm ² , about 3 cm ² to about 16 cm ² , about 3 cm ² to about 14 cm ² , about 3 cm ² to about 12 cm ² , about 3 cm ² to about 10 cm ² , about 3 cm ² to about 8 cm ² , about 3 cm ² to about 6 cm ² , about 3 cm ² to about 5 cm ² , about 3 cm ² to about 4 cm ² ,

about 4 cm ² to about 60 cm ² , about 4 cm ² to about 58 cm ² , about 4 cm ² to about 56 cm ² , about 4 cm ² to about 54 cm ² , about 4 cm ² to about 52 cm ² , about 4 cm ² to about 50 cm ² , about 4 cm ² to about 48 cm ² , about 4 cm ² to about 46 cm ² , about 4 cm ² to about 44 cm ² , about 4 cm ² to about 42 cm ² , about 4 cm ² to about 40 cm ² , about 4 cm ² to about 38 cm ² , about 4 cm ² to about 36 cm ² , about 4 cm ² to about 34 cm ² , about 4 cm ² to about 32 cm ² , about 4 cm ² to about 30 cm ² , about 4 cm ² to about 28 cm ² , about 4 cm ² to about 26 cm ² , about 4 cm ² to about 24 cm ² , about 4 cm ² to about 22 cm ² , about 4 cm ² to about 20 cm ² , about 4 cm ² to about 18 cm ² , about 4 cm ² to about 16 cm ² , about 4 cm ² to about 14 cm ² , about 4 cm ² to about 12 cm ² , about 4 cm ² to about 10 cm ² , about 4 cm ² to about 8 cm ² , about 4 cm ² to about 6 cm ² , about 4 cm ² to about 5 cm ² , about 5 cm ² to about 60 cm ² , about 5 cm ² to about 58 cm ² , about 5 cm ² to about 56 cm ² , about 5 cm ² to about 54 cm ² , about 5 cm ² to about 52 cm ² , about 5 cm ² to about 50 cm ² , about 5 cm ² to about 48 cm ² , about 5 cm ² to about 46 cm ² , about 5 cm ² to about 44 cm ² , about 5 cm ² to about 42 cm ² , about 5 cm ² to about 40 cm ² , about 5 cm ² to about 38 cm ² , about 5 cm ² to about 36 cm ² , about 5 cm ² to about 34 cm ² , about 5 cm ² to about 32 cm ² , about 5 cm ² to about 30 cm ² , about 5 cm ² to about 28 cm ² , about 5 cm ² to about 26 cm ² , about 5 cm ² to about 24 cm ² , about 5 cm ² to about 22 cm ² , about 5 cm ² to about 20 cm ² , about 5 cm ² to about 18 cm ² , about 5 cm ² to about 16 cm ² , about 5 cm ² to about 14 cm ² , about 5 cm ² to about 12 cm ² , about 5 cm ² to about 10 cm ² , about 5 cm ² to about 8 cm ² , about 5 cm ² to about 6 cm ² ,

about 6 cm ² to about 60 cm ² , about 6 cm ² to about 58 cm ² , about 6 cm ² to about 56 cm ² , about 6 cm ² to about 54 cm ² , about 6 cm ² to about 52 cm ² , about 6 cm ² to about 50 cm ² , about 6 cm ² to about 48 cm ² , about 6 cm ² to about 46 cm ² , about 6 cm ² to about 44 cm ² , about 6 cm ² to about 42 cm ² , about 6 cm ² to about 40 cm ² , about 6 cm ² to about 38 cm ² , about 6 cm ² to about 36 cm ² , about 6 cm ² to about 34 cm ² , about 6 cm ² to about 32 cm ² , about 6 cm ² to about 30 cm ² , about 6 cm ² to about 28 cm ² , about 6 cm ² to about 26 cm ² , about 6 cm ² to about 24 cm ² , about 6 cm ² to about 22 cm ² , about 6 cm ² to about 20 cm ² , about 6 cm ² to about 18 cm ² , about 6 cm ² to about 16 cm ² , about 6 cm ² to about 14 cm ² , about 6 cm ² to about 12 cm ² , about 6 cm ² to about 10 cm ² , about 6 cm ² to about 8 cm ² ,

about 8 cm ² to about 60 cm ² , about 8 cm ² to about 58 cm ² , about 8 cm ² to about 56 cm ² , about 8 cm ² to about 54 cm ² , about 8 cm ² to about 52 cm ² , about 8 cm ² to about 50 cm ² , about 8 cm ² to about 48 cm ² , about 8 cm ² to about 46 cm ² , about 8 cm ² to about 44 cm ² , about 8 cm ² to about 42 cm ² , about 8 cm ² to about 40 cm ² , about 8 cm ² to about 38 cm ² , about 8 cm ² to about 36 cm ² , about 8 cm ² to about 34 cm ² , about 8 cm ² to about 32 cm ² ,

about 8 cm ² to about 30 cm ² , about 8 cm ² to about 28 cm ² , about 8 cm ² to about 26 cm ² , about 8 cm ² to about 24 cm ² , about 8 cm ² to about 22 cm ² , about 8 cm ² to about 20 cm ² , about 8 cm ² to about 18 cm ² , about 8 cm ² to about 16 cm ² , about 8 cm ² to about 14 cm ² , about 8 cm ² to about 12 cm ² , about 8 cm ² to about 10 cm ² , about 10 cm ² to about 60 cm ² , about 10 cm ² to about 58 cm ² , about 10 cm ² to about 56 cm ² , about 10 cm ² to about 54 cm ² , about 10 cm ² to about 52 cm ² , about 10 cm ² to about 50 cm ² , about 10 cm ² to about 48 cm ² , about 10 cm ² to about 46 cm ² , about 10 cm ² to about 44 cm ² , about 10 cm ² to about 42 cm ² , about 10 cm ² to about 40 cm ² , about 10 cm ² to about 38 cm ² , about 10 cm ² to about 36 cm ² , about 10 cm ² to about 34 cm ² , about 10 cm ² to about 32 cm ² , about 10 cm ² to about 30 cm ² , about 10 cm ² to about 28 cm ² , about 10 cm ² to about 26 cm ² , about 10 cm ² to about 24 cm ² , about 10 cm ² to about 22 cm ² , about 10 cm ² to about 20 cm ² , about 10 cm ² to about 18 cm ² , about 10 cm ² to about 16 cm ² , about 10 cm ² to about 14 cm ² , about 10 cm ² to about 12 cm ² , about 12 cm ² to about 60 cm ² , about 12 cm ² to about 58 cm ² , about 12 cm ² to about 56 cm ² , about 12 cm ² to about 54 cm ² , about 12 cm ² to about 52 cm ² , about 12 cm ² to about 50 cm ² , about 12 cm ² to about 48 cm ² , about 12 cm ² to about 46 cm ² , about 12 cm ² to about 44 cm ² , about 12 cm ² to about 42 cm ² , about 12 cm ² to about 40 cm ² , about 12 cm ² to about 38 cm ² , about 12 cm ² to about 36 cm ² , about 12 cm ² to about 34 cm ² , about 12 cm ² to about 32 cm ² , about 12 cm ² to about 30 cm ² , about 12 cm ² to about 28 cm ² , about 12 cm ² to about 26 cm ² , about 12 cm ² to about 24 cm ² , about 12 cm ² to about 22 cm ² , about 12 cm ² to about 20 cm ² , about 12 cm ² to about 18 cm ² , about 12 cm ² to about 16 cm ² , about 12 cm ² to about 14 cm ² , about 14 cm ² to about 60 cm ² , about 14 cm ² to about 58 cm ² , about 14 cm ² to about 56 cm ² , about 14 cm ² to about 54 cm ² , about 14 cm ² to about 52 cm ² , about 14 cm ² to about 50 cm ² , about 14 cm ² to about 48 cm ² , about 14 cm ² to about 46 cm ² , about 14 cm ² to about 44 cm ² , about 14 cm ² to about 42 cm ² , about 14 cm ² to about 40 cm ² , about 14 cm ² to about 38 cm ² , about 14 cm ² to about 36 cm ² , about 14 cm ² to about 34 cm ² , about 14 cm ² to about 32 cm ² , about 14 cm ² to about 30 cm ² , about 14 cm ² to about 28 cm ² , about 14 cm ² to about 26 cm ² , about 14 cm ² to about 24 cm ² , about 14 cm ² to about 22 cm ² , about 14 cm ² to about 20 cm ² , about 14 cm ² to about 18 cm ² , about 14 cm ² to about 16 cm ² , about 16 cm ² to about 60 cm ² , about 16 cm ² to about 58 cm ² , about 16 cm ² to about 56 cm ² , about 16 cm ² to about 54 cm ² , about 16 cm ² to about 52 cm ² , about 16 cm ² to about 50 cm ² , about 16 cm ² to about 48 cm ² , about 16 cm ² to about 46 cm ² , about 16 cm ² to about 44 cm ² , about 16 cm ² to about 42 cm ² , about 16 cm ² to about 40 cm ² , about 16 cm ² to about 38 cm ² , about 16 cm ² to about 36 cm ² , about 16 cm ² to about 34 cm ² , about 16 cm ² to about 32 cm ² , about 16 cm ² to about 30 cm ² , about 16 cm ² to about 28 cm ² , about 16 cm ² to about 26 cm ² , about 16 cm ² to about 24 cm ² , about 16 cm ² to about 22 cm ² , about 16 cm ² to about 20 cm ² , about 16 cm ² to about 18 cm ² , about 18 cm ² to about 60 cm ² , about 18 cm ² to about 58 cm ² , about 18 cm ² to about 56 cm ² , about 18 cm ² to about 54 cm ² , about 18 cm ² to about 52 cm ² , about 18 cm ² to about 50 cm ² , about 18 cm ² to about 48 cm ² , about 18 cm ² to about 46 cm ² , about 18 cm ² to about 44 cm ² , about 18 cm ² to about 42 cm ² , about 18 cm ² to about 40 cm ² , about 18 cm ² to about 38 cm ² , about 18 cm ² to about 36 cm ² , about 18 cm ² to about 34 cm ² , about 18 cm ² to about 32 cm ² , about 18 cm ² to about 30 cm ² , about 18 cm ² to about 28 cm ² , about 18 cm ² to about 26 cm ² , about 18 cm ² to about 24 cm ² , about 18 cm ² to about 22 cm ² , about 18 cm ² to about 20 cm ² , about 20 cm ² to about 60 cm ² , about 20 cm ² to about 58 cm ² , about 20 cm ² to about 56 cm ² , about 20 cm ² to about 54 cm ² , about 20 cm ² to about 52 cm ² , about 20 cm ² to about 50 cm ² , about 20 cm ² to about 48 cm ² , about 20 cm ² to about 46 cm ² , about 20 cm ² to about 44 cm ² , about 20 cm ² to about 42 cm ² , about 20 cm ² to about 40 cm ² , about 20 cm ² to about 38 cm ² , about 20 cm ² to about 36 cm ² , about 20 cm ² to about 34 cm ² , about 20 cm ² to about 32 cm ² , about 20 cm ² to about 30 cm ² , about 20 cm ² to about 28 cm ² , about 20 cm ² to about 26 cm ² , about 20 cm ² to about 24 cm ² , about 20 cm ² to about 22 cm ² , about 22 cm ² to about 60 cm ² , about 22 cm ² to about 58 cm ² , about 22 cm ² to about 56 cm ² , about 22 cm ² to about 54 cm ² , about 22 cm ² to about 52 cm ² , about 22 cm ² to about 50 cm ² , about 22 cm ² to about 48 cm ² , about 22 cm ² to about 46 cm ² , about 22 cm ² to about 44 cm ² , about 22 cm ² to about 42 cm ² , about 22 cm ² to about 40 cm ² , about 22 cm ² to about 38 cm ² , about 22 cm ² to about 36 cm ² , about 22 cm ² to about 34 cm ² , about 22 cm ² to about 32 cm ² , about 22 cm ² to about 30 cm ² , about 22 cm ² to about 28 cm ² , about 22 cm ² to about 26 cm ² , about 22 cm ² to about 24 cm ² , about 24 cm ² to about 60 cm ² , about 24 cm ² to about 58 cm ² , about 24 cm ² to about 56 cm ² , about 24 cm ² to about 54 cm ² , about 24 cm ² to about 52 cm ² , about 24 cm ² to about 50 cm ² , about 24 cm ² to about 48 cm ² , about 24 cm ² to about 46 cm ² , about 24 cm ² to about 44 cm ² , about 24 cm ² to about 42 cm ² , about 24 cm ² to about 40 cm ² , about 24 cm ² to about 38 cm ² , about 24 cm ² to about 36 cm ² , about 24 cm ² to about 34 cm ² , about 24 cm ² to about 32 cm ² , about 24 cm ² to about 30 cm ² , about 24 cm ² to about 28 cm ² , about 24 cm ² to about 26 cm ² ,

about 26 cm ² to about 60 cm ² , about 26 cm ² to about 58 cm ² , about 26 cm ² to about 56 cm ² , about 26 cm ² to about 54 cm ² , about 26 cm ² to about 52 cm ² , about 26 cm ² to about 50 cm ² , about 26 cm ² to about 48 cm ² , about 26 cm ² to about 46 cm ² , about 26 cm ² to about 44 cm ² , about 26 cm ² to about 42 cm ² , about 26 cm ² to about 40 cm ² , about 26 cm ² to about 38 cm ² , about 26 cm ² to about 36 cm ² , about 26 cm ² to about 34 cm ² , about 26 cm ² to about 32 cm ² , about 26 cm ² to about 30 cm ² , about 26 cm ² to about 28 cm ² ,

about 28 cm ² to about 60 cm ² , about 28 cm ² to about 58 cm ² , about 28 cm ² to about 56 cm ² , about 28 cm ² to about 54 cm ² , about 28 cm ² to about 52 cm ² , about 28 cm ² to about 50 cm ² , about 28 cm ² to about 48 cm ² , about 28 cm ² to about 46 cm ² , about 28 cm ² to about 44 cm ² , about 28 cm ² to about 42 cm ² , about 28 cm ² to about 40 cm ² , about 28 cm ² to about 38 cm ² , about 28 cm ² to about 36 cm ² , about 28 cm ² to about 34 cm ² , about 28 cm ² to about 32 cm ² , about 28 cm ² to about 30 cm ² , about 30 cm ² to about 60 cm ² , about 30 cm ² to about 58 cm ² , about 30 cm ² to about 56 cm ² , about 30 cm ² to about 54 cm ² , about 30 cm ² to about 52 cm ² , about 30 cm ² to about 50 cm ² , about 30 cm ² to about 48 cm ² , about 30 cm ² to about 46 cm ² , about 30 cm ² to about 44 cm ² , about 30 cm ² to about 42 cm ² , about 30 cm ² to about 40 cm ² , about 30 cm ² to about 38 cm ² , about 30 cm ² to about 36 cm ² , about 30 cm ² to about 34 cm ² , about 30 cm ² to about 32 cm ² , about 32 cm ² to about 60 cm ² , about 32 cm ² to about 58 cm ² , about 32 cm ² to about 56 cm ² , about 32 cm ² to about 54 cm ² , about 32 cm ² to about 52 cm ² , about 32 cm ² to about 50 cm ² , about 32 cm ² to about 48 cm ² , about 32 cm ² to about 46 cm ² , about 32 cm ² to about 44 cm ² , about 32 cm ² to about 42 cm ² , about 32 cm ² to about 40 cm ² , about 32 cm ² to about 38 cm ² , about 32 cm ² to about 36 cm ² , about 32 cm ² to about 34 cm ² , about 34 cm ² to about 60 cm ² , about 34 cm ² to about 58 cm ² , about 34 cm ² to about 56 cm ² , about 34 cm ² to about 54 cm ² , about 34 cm ² to about 52 cm ² , about 34 cm ² to about 50 cm ² , about 34 cm ² to about 48 cm ² , about 34 cm ² to about 46 cm ² , about 34 cm ² to about 44 cm ² , about 34 cm ² to about 42 cm ² , about 34 cm ² to about 40 cm ² , about 34 cm ² to about 38 cm ² , about 34 cm ² to about 36 cm ² , about 36 cm ² to about 60 cm ² , about 36 cm ² to about 58 cm ² , about 36 cm ² to about 56 cm ² , about 36 cm ² to about 54 cm ² , about 36 cm ² to about 52 cm ² , about 36 cm ² to about 50 cm ² , about 36 cm ² to about 48 cm ² , about 36 cm ² to about 46 cm ² , about 36 cm ² to about 44 cm ² , about 36 cm ² to about 42 cm ² , about 36 cm ² to about 40 cm ² , about 36 cm ² to about 38 cm ² , about 38 cm ² to about 60 cm ² , about 38 cm ² to about 58 cm ² , about 38 cm ² to about 56 cm ² , about 38 cm ² to about 54 cm ² , about 38 cm ² to about 52 cm ² , about 38 cm ² to about 50 cm ² , about 38 cm ² to about 48 cm ² , about 38 cm ² to about 46 cm ² , about 38 cm ² to about 44 cm ² , about 38 cm ² to about 42 cm ² , about 38 cm ² to about 40 cm ² , about 40 cm ² to about 60 cm ² , about 40 cm ² to about 58 cm ² , about 40 cm ² to about 56 cm ² , about 40 cm ² to about 54 cm ² , about 40 cm ² to about 52 cm ² , about 40 cm ² to about 50 cm ² , about 40 cm ² to about 48 cm ² , about 40 cm ² to about 46 cm ² , about 40 cm ² to about 44 cm ² , about 40 cm ² to about 42 cm ² , about 42 cm ² to about 60 cm ² , about 42 cm ² to about 58 cm ² , about 42 cm ² to about 56 cm ² , about 42 cm ² to about 54 cm ² , about 42 cm ² to about 52 cm ² , about 42 cm ² to about 50 cm ² , about 42 cm ² to about 48 cm ² , about 42 cm ² to about 46 cm ² , about 42 cm ² to about 44 cm ² , about 44 cm ² to about 60 cm ² , about 44 cm ² to about 58 cm ² , about 44 cm ² to about 56 cm ² , about 44 cm ² to about 54 cm ² , about 44 cm ² to about 52 cm ² , about 44 cm ² to about 50 cm ² , about 44 cm ² to about 48 cm ² , about 44 cm ² to about 46 cm ² , about 46 cm ² to about 60 cm ² , about 46 cm ² to about 58 cm ² , about 46 cm ² to about 56 cm ² , about 46 cm ² to about 54 cm ² , about 46 cm ² to about 52 cm ² , about 46 cm ² to about 50 cm ² , about 46 cm ² to about 48 cm ² , about 48 cm ² to about 60 cm ² , about 48 cm ² to about 58 cm ² , about 48 cm ² to about 56 cm ² , about 48 cm ² to about 54 cm ² , about 48 cm ² to about 52 cm ² , about 48 cm ² to about 50 cm ² , about 50 cm ² to about 60 cm ² , about 50 cm ² to about 58 cm ² , about 50 cm ² to about 56 cm ² , about 50 cm ² to about 54 cm ² , about 50 cm ² to about 52 cm ² , about 52 cm ² to about 60 cm ² , about 52 cm ² to about 58 cm ² , about 52 cm ² to about 56 cm ² , about 52cm ² to about 54 cm ² , about 54 cm ² to about 60 cm ² , about 54 cm ² to about 58 cm ² , about 54 cm ² to about 56 cm ² , about 56 cm ² to about 60 cm ² , about 56 cm ² to about 58 cm ² , or about 58 cm ² to about 60 cm ² .

In some embodiments of any of the pods described herein, the total sum of the surface area of the least one areas covered by the semi-porous or porous material is about 0.5 mm ² to about 150 cm ² , about 0.5 mm ² to about 140 cm ² , about 0.5 mm ² to about 130 cm ² , about 0.5 mm ² to about 120 cm ² , about 0.5 mm ² to about 110 cm ² , about 0.5 mm ² to about 100 cm ² , about 0.5 mm ² to about 90 cm ² , about 0.5 mm ² to about 80 cm ² , about 0.5 mm ² to about 70 cm ² , about 0.5 mm ² to about 60 cm ² , about 0.5 mm ² to about 50 cm ² , about 0.5 mm ² to about 40 cm ² , about 0.5 mm ² to about 30 cm ² , about 0.5 mm ² to about 20 cm ² , about 0.5 mm ² to about 18 cm ² , about 0.5 mm ² to about 16 cm ² , about 0.5 mm ² to about 14 cm ² , about 0.5 mm ² to about 12 cm ² , about 0.5 mm ² to about 10 cm ² , about 0.5 mm ² to about 8 cm ² , about 0.5 mm ² to about 6 cm ² , about 0.5 mm ² to about 5 cm ² , about 0.5 mm ² to about 4 cm ² , about 0.5 mm ² to about 3 cm ² , about 0.5 mm ² to about 2 cm ² , about 0.5 mm ² to about 1 cm ² , about 0.5 mm ² to about 5 mm ² , about 5 mm ² to about 150 cm ² , about 5 mm ² to about 140 cm ² , about 5 mm ² to about 130 cm ² , about 5 mm ² to about 120 cm ² , about 5 mm ² to about 110 cm ² , about 5 mm ² to about 100 cm ² , about 5 mm ² to about 90 cm ² , about 5 mm ² to about 80 cm ² , about 5 mm ² to about 70 cm ² , about 5 mm ² to about 60 cm ² , about 5 mm ² to about 50 cm ² , about 5 mm ² to about 40 cm ² , about 5 mm ² to about 30 cm ² , about 5 mm ² to about 20 cm ² , about 5 mm ² to about 18 cm ² , about 5 mm ² to about 16 cm ² , about 5 mm ² to about 14 cm ² , about 5 mm ² to about 12 cm ² , about 5 mm ² to about 10 cm ² , about 5 mm ² to about 8 cm ² , about 5 mm ² to about 6 cm ² , about 5 mm ² to about 5 cm ² , about 5 mm ² to about 4 cm ² , about 5 mm ² to about 3 cm ² , about 5 mm ² to about 2 cm ² , about 5 mm ² to about 1 cm ² , about 1 cm ² to about 150 cm ² , about 1 cm ² to about 140 cm ² , about 1 cm ² to about 130 cm ² , about 1 cm ² to about 120 cm ² , about 1 cm ² to about 110 cm ² , about 1 cm ² to about 100 cm ² , about 1 cm ² to about 90 cm ² , about 1 cm ² to about 80 cm ² , about 1 cm ² to about 70 cm ² , about 1 cm ² to about 60 cm ² , about 1 cm ² to about 50 cm ² , about 1 cm ² to about 40 cm ² , about 1 cm ² to about 30 cm ² , about 1 cm ² to about 20 cm ² , about 1 cm ² to about 18 cm ² , about 1 cm ² to about 16 cm ² , about 1 cm ² to about 14 cm ² , about 1 cm ² to about 12 cm ² , about 1 cm ² to about 10 cm ² , about 1 cm ² to about 8 cm ² , about 1 cm ² to about 6 cm ² , about 1 cm ² to about 5 cm ² , about 1 cm ² to about 4 cm ² , about 1 cm ² to about 3 cm ² , about 1 cm ² to about 2 cm ² , about 2 cm ² to about 150 cm ² , about 2 cm ² to about 140 cm ² , about 2 cm ² to about 130 cm ² , about 2 cm ² to about 120 cm ² , about 2 cm ² to about 110 cm ² , about 2 cm ² to about 100 cm ² , about 2 cm ² to about 90 cm ² , about 2 cm ² to about 80 cm ² , about 2 cm ² to about 70 cm ² , about 2 cm ² to about 60 cm ² , about 2 cm ² to about 50 cm ² , about 2 cm ² to about 40 cm ² , about 2 cm ² to about 30 cm ² , about 2 cm ² to about 20 cm ² , about 2 cm ² to about 18 cm ² , about 2 cm ² to about 16 cm ² , about 2 cm ² to about 14 cm ² , about 2 cm ² to about 12 cm ² , about 2 cm ² to about 10 cm ² , about 2 cm ² to about 8 cm ² , about 2 cm ² to about 6 cm ² , about 2 cm ² to about 5 cm ² , about 2 cm ² to about 4 cm ² , about 2 cm ² to about 3 cm ² , about 3 cm ² to about 150 cm ² , about 3 cm ² to about 140 cm ² , about 3 cm ² to about 130 cm ² , about 3 cm ² to about 120 cm ² , about 3 cm ² to about 110 cm ² , about 3 cm ² to about 100 cm ² , about 3 cm ² to about 90 cm ² , about 3 cm ² to about 80 cm ² , about 3 cm ² to about 70 cm ² , about 3 cm ² to about 60 cm ² , about 3 cm ² to about 50 cm ² , about 3 cm ² to about 40 cm ² , about 3 cm ² to about 30 cm ² , about 3 cm ² to about 20 cm ² , about 3 cm ² to about 18 cm ² , about 3 cm ² to about 16 cm ² , about 3 cm ² to about 14 cm ² , about 3 cm ² to about 12 cm ² , about 3 cm ² to about 10 cm ² , about 3 cm ² to about 8 cm ² , about 3 cm ² to about 6 cm ² , about 3 cm ² to about 5 cm ² , about 3 cm ² to about 4 cm ² , about 4 cm ² to about 150 cm ² , about 4 cm ² to about 140 cm ² , about 4 cm ² to about 130 cm ² , about 4 cm ² to about 120 cm ² , about 4 cm ² to about 110 cm ² , about 4 cm ² to about 100 cm ² , about 4 cm ² to about 90 cm ² , about 4 cm ² to about 80 cm ² , about 4 cm ² to about 70 cm ² , about 4 cm ² to about 60 cm ² , about 4 cm ² to about 50 cm ² , about 4 cm ² to about 40 cm ² , about 4 cm ² to about 30 cm ² , about 4 cm ² to about 20 cm ² , about 4 cm ² to about 18 cm ² , about 4 cm ² to about 16 cm ² , about 4 cm ² to about 14 cm ² , about 4 cm ² to about 12 cm ² , about 4 cm ² to about 10 cm ² , about 4 cm ² to about 8 cm ² , about 4 cm ² to about 6 cm ² , about 4 cm ² to about 5 cm ² , about 5 cm ² to about 150 cm ² , about 5 cm ² to about 140 cm ² , about 5 cm ² to about 130 cm ² , about 5 cm ² to about 120 cm ² , about 5 cm ² to about 110 cm ² , about 5 cm ² to about 100 cm ² , about 5 cm ² to about 90 cm ² , about 5 cm ² to about 80 cm ² , about 5 cm ² to about 70 cm ² , about 5 cm ² to about 60 cm ² , about 5 cm ² to about 50 cm ² , about 5 cm ² to about 40 cm ² , about 5 cm ² to about 30 cm ² , about 5 cm ² to about 20 cm ² , about 5 cm ² to about 18 cm ² , about 5 cm ² to about 16 cm ² , about 5 cm ² to about 14 cm ² , about 5 cm ² to about 12 cm ² , about 5 cm ² to about 10 cm ² , about 5 cm ² to about 8 cm ² , about 5 cm ² to about 6 cm ² , about 6 cm ² to about 150 cm ² , about 6 cm ² to about 140 cm ² , about 6 cm ² to about 130 cm ² , about 6 cm ² to about 120 cm ² , about 6 cm ² to about 110 cm ² , about 6 cm ² to about 100 cm ² , about 6 cm ² to about 90 cm ² , about 6 cm ² to about 80 cm ² , about 6 cm ² to about 70 cm ² , about 6 cm ² to about 60 cm ² , about 6 cm ² to about 50 cm ² , about 6 cm ² to about 40 cm ² , about 6 cm ² to about 30 cm ² , about 6 cm ² to about 20 cm ² , about 6 cm ² to about 18 cm ² , about 6 cm ² to about 16 cm ² , about 6 cm ² to about 14 cm ² , about 6 cm ² to about 12 cm ² , about 6 cm ² to about 10 cm ² , about 6 cm ² to about 8 cm ² , about 8 cm ² to about 150 cm ² , about 8 cm ² to about 140 cm ² , about 8 cm ² to about 130 cm ² , about 8 cm ² to about 120 cm ² , about 8 cm ² to about 110 cm ² , about 8 cm ² to about 100 cm ² , about 8 cm ² to about 90 cm ² , about 8 cm ² to about 80 cm ² , about 8 cm ² to about 70 cm ² , about 8 cm ² to about 60 cm ² , about 8 cm ² to about 50 cm ² , about 8 cm ² to about 40 cm ² , about 8 cm ² to about 30 cm ² , about 8 cm ² to about 20 cm ² , about 8 cm ² to about 18 cm ² , about 8 cm ² to about 16 cm ² , about 8 cm ² to about 14 cm ² , about 8 cm ² to about 12 cm ² , about 8 cm ² to about 10 cm ² , about 10 cm ² to about 150 cm ² , about 10 cm ² to about 140 cm ² , about 10 cm ² to about 130 cm ² , about 10 cm ² to about 120 cm ² , about 10 cm ² to about 110 cm ² , about 10 cm ² to about 100 cm ² , about 10 cm ² to about 90 cm ² , about 10 cm ² to about 80 cm ² , about 10 cm ² to about 70 cm ² , about 10 cm ² to about 60 cm ² , about 10 cm ² to about 50 cm ² , about 10 cm ² to about 40 cm ² , about 10 cm ² to about 30 cm ² , about 10 cm ² to about 20 cm ² , about 10 cm ² to about 18 cm ² , about 10 cm ² to about 16 cm ² , about 10 cm ² to about 14 cm ² , about 10 cm ² to about 12 cm ² , about 12 cm ² to about 150 cm ² , about 12 cm ² to about 140 cm ² , about 12 cm ² to about 130 cm ² , about 12 cm ² to about 120 cm ² , about 12 cm ² to about 110 cm ² , about 12 cm ² to about 100 cm ² , about 12 cm ² to about 90 cm ² , about 12 cm ² to about 80 cm ² , about 12 cm ² to about 70 cm ² , about 12 cm ² to about 60 cm ² , about 12 cm ² to about 50 cm ² , about 12 cm ² to about 40 cm ² , about 12 cm ² to about 30 cm ² , about 12 cm ² to about 20 cm ² , about 12 cm ² to about 18 cm ² , about 12 cm ² to about 16 cm ² , about 12 cm ² to about 14 cm ² , about 14 cm ² to about 150 cm ² , about 14 cm ² to about 140 cm ² , about 14 cm ² to about 130 cm ² , about 14 cm ² to about 120 cm ² , about 14 cm ² to about 110 cm ² , about 14 cm ² to about 100 cm ² , about 14 cm ² to about 90 cm ² , about 14 cm ² to about 80 cm ² , about 14 cm ² to about 70 cm ² , about 14 cm ² to about 60 cm ² , about 14 cm ² to about 50 cm ² , about 14 cm ² to about 40 cm ² , about 14 cm ² to about 30 cm ² , about 14 cm ² to about 20 cm ² , about 14 cm ² to about 18 cm ² , about 14 cm ² to about 16 cm ² , about 16 cm ² to about 150 cm ² , about 16 cm ² to about 140 cm ² , about 16 cm ² to about 130 cm ² , about 16 cm ² to about 120 cm ² , about 16 cm ² to about 110 cm ² , about 16 cm ² to about 100 cm ² , about 16 cm ² to about 90 cm ² , about 16 cm ² to about 80 cm ² , about 16 cm ² to about 70 cm ² , about 16 cm ² to about 60 cm ² , about 16 cm ² to about 50 cm ² , about 16 cm ² to about 40 cm ² , about 16 cm ² to about 30 cm ² , about 16 cm ² to about 20 cm ² , about 16 cm ² to about 18 cm ² , about 18 cm ² to about 150 cm ² , about 18 cm ² to about 140 cm ² , about 18 cm ² to about 130 cm ² , about 18 cm ² to about 120 cm ² , about 18 cm ² to about 110 cm ² , about 18 cm ² to about 100 cm ² , about 18 cm ² to about 90 cm ² , about 18 cm ² to about 80 cm ² , about 18 cm ² to about 70 cm ² , about 18 cm ² to about 60 cm ² , about 18 cm ² to about 50 cm ² , about 18 cm ² to about 40 cm ² , about 18 cm ² to about 30 cm ² , about 18 cm ² to about 20 cm ² , about 20 cm ² to about 150 cm ² , about 20 cm ² to about 140 cm ² , about 20 cm ² to about 130 cm ² , about 20 cm ² to about 120 cm ² , about 20 cm ² to about 110 cm ² , about 20 cm ² to about 100 cm ² , about 20 cm ² to about 90 cm ² , about 20 cm ² to about 80 cm ² , about 20 cm ² to about 70 cm ² , about 20 cm ² to about 60 cm ² , about 20 cm ² to about 50 cm ² , about 20 cm ² to about 40 cm ² , about 20 cm ² to about 30 cm ² , about 30 cm ² to about 150 cm ² , about 30 cm ² to about 140 cm ² , about 30 cm ² to about 130 cm ² , about 30 cm ² to about 120 cm ² , about 30 cm ² to about 110 cm ² , about 30 cm ² to about 100 cm ² , about 30 cm ² to about 90 cm ² , about 30 cm ² to about 80 cm ² , about 30 cm ² to about 70 cm ² , about 30 cm ² to about 60 cm ² , about 30 cm ² to about 50 cm ² , about 30 cm ² to about 40 cm ² , about 40 cm ² to about 150 cm ² , about 40 cm ² to about 140 cm ² , about 40 cm ² to about 130 cm ² , about 40 cm ² to about 120 cm ² , about 40 cm ² to about 110 cm ² , about 40 cm ² to about 100 cm ² , about 40 cm ² to about 90 cm ² , about 40 cm ² to about 80 cm ² , about 40 cm ² to about 70 cm ² , about 40 cm ² to about 60 cm ² , about 40 cm ² to about 50 cm ² , about 50 cm ² to about 150 cm ² , about 50 cm ² to about 140 cm ² , about 50 cm ² to about 130 cm ² , about 50 cm ² to about 120 cm ² , about 50 cm ² to about 110 cm ² , about 50 cm ² to about 100 cm ² , about 50 cm ² to about 90 cm ² , about 50 cm ² to about 80 cm ² , about 50 cm ² to about 70 cm ² , about 50 cm ² to about 60 cm ² , about 60 cm ² to about 150 cm ² , about 60 cm ² to about 140 cm ² , about 60 cm ² to about 130 cm ² , about 60 cm ² to about 120 cm ² , about 60 cm ² to about 110 cm ² , about 60 cm ² to about 100 cm ² , about 60 cm ² to about 90 cm ² , about 60 cm ² to about 80 cm ² , about 60 cm ² to about 70 cm ² , about 70 cm ² to about 150 cm ² , about 70 cm ² to about 140 cm ² , about 70 cm ² to about 130 cm ² , about 70 cm ² to about 120 cm ² , about 70 cm ² to about 110 cm ² , about 70 cm ² to about 100 cm ² , about 70 cm ² to about 90 cm ² , about 70 cm ² to about 80 cm ² , about 80 cm ² to about 150 cm ² , about 80 cm ² to about 140 cm ² , about 80 cm ² to about 130 cm ² , about 80 cm ² to about 120 cm ² , about 80 cm ² to about 110 cm ² , about 80 cm ² to about 100 cm ² , about 80 cm ² to about 90 cm ² , about 90 cm ² to about 150 cm ² , about 90 cm ² to about 140 cm ² , about 90 cm ² to about 130 cm ² , about 90 cm ² to about 120 cm ² , about 90 cm ² to about 110 cm ² , about 90 cm ² to about 100 cm ² , about 100 cm ² to about 150 cm ² , about 100 cm ² to about 140 cm ² , about 100 cm ² to about 130 cm ² , about 100 cm ² to about 120 cm ² , about 100 cm ² to about 110 cm ² , about 110 cm ² to about 150 cm ² , about 110 cm ² to about 140 cm ² , about 110 cm ² to about 130 cm ² , about 110 cm ² to about 120 cm ² , about 120 cm ² to about 150 cm ² , about 120 cm ² to about 140 cm ² , about 120 cm ² to about 130 cm ² , about 130 cm ² to about 150 cm ² , about 130 cm ² to about 140 cm ² , or about 140 cm ² to about 150 cm ² .

In some embodiments of any of the pods described herein, where the wall comprises at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, the percentage of the total surface area of the wall that is represented by at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod is about 0.1% to about 95%, about 0.1% to about 90%, about 0.1% to about 85%, about 0.1% to about 80%, about 0.1% to about 75%, about 0.1% to about 70%, about 0.1% to about 65%, about 0.1% to about 60%, about 0.1% to about 55%, about 0.1% to about 50%, about 0.1% to about 45%, about 0.1% to about 40%, about 0.1% to about 35%, about 0.1% to about 30%, about 0.1% to about 25%, about 0.1% to about 20%, about 0.1% to about 18%, about 0.1% to about 16%, about 0.1% to about 14%, about 0.1% to about 12%, about 0.1% to about 10%, about 0.1% to about 8%, about 0.1% to about 6%, about 0.1% to about 5%, about 0.1% to about 4%, about 0.1% to about 3%, about 0.1% to about 2%, about 0.1% to about 1%, about 0.1% to about 0.5%, about 0.1% to about 0.2%, about 0.2% to about 95%, about 0.2% to about 90%, about 0.2% to about 85%, about 0.2% to about 80%, about 0.2% to about 75%, about 0.2% to about 70%, about 0.2% to about 65%, about 0.2% to about 60%, about 0.2% to about 55%, about 0.2% to about 50%, about 0.2% to about 45%, about 0.2% to about 40%, about 0.2% to about 35%, about 0.2% to about 30%, about 0.2% to about 25%, about 0.2% to about 20%, about 0.2% to about 18%, about 0.2% to about 16%, about 0.2% to about 14%, about 0.2% to about 12%, about 0.2% to about 10%, about 0.2% to about 8%, about 0.2% to about 6%, about 0.2% to about 5%, about 0.2% to about 4%, about 0.2% to about 3%, about 0.2% to about 2%, about 0.2% to about 1%, about 0.2% to about 0.5%, about 0.5% to about 95%, about 0.5% to about 90%, about 0.5% to about 85%, about 0.5% to about 80%, about 0.5% to about 75%, about 0.5% to about 70%, about 0.5% to about 65%, about 0.5% to about 60%, about 0.5% to about 55%, about 0.5% to about 50%, about 0.5% to about 45%, about 0.5% to about 40%, about 0.5% to about 35%, about 0.5% to about 30%, about 0.5% to about 25%, about 0.5% to about 20%, about 0.5% to about 18%, about 0.5% to about 16%, about 0.5% to about 14%, about 0.5% to about 12%, about 0.5% to about 10%, about 0.5% to about 8%, about 0.5% to about 6%, about 0.5% to about 5%, about 0.5% to about 4%, about 0.5% to about 3%, about 0.5% to about 2%, about 0.5% to about 1%, about 1% to about 95%, about 1% to about 90%, about 1% to about 85%, about 1% to about 80%, about 1% to about 75%, about 1% to about 70%, about 1% to about 65%, about 1% to about 60%, about 1% to about 55%, about 1% to about 50%, about 1% to about 45%, about 1% to about 40%, about 1% to about 35%, about 1% to about 30%, about 1% to about 25%, about 1% to about 20%, about 1% to about 18%, about 1% to about 16%, about 1% to about 14%, about 1% to about 12%, about 1% to about 10%, about 1% to about 8%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 95%, about 2% to about 90%, about 2% to about 85%, about 2% to about 80%, about 2% to about 75%, about 2% to about 70%, about 2% to about 65%, about 2% to about 60%, about 2% to about 55%, about 2% to about 50%, about 2% to about 45%, about 2% to about 40%, about 2% to about 35%, about 2% to about 30%, about 2% to about 25%, about 2% to about 20%, about 2% to about 18%, about 2% to about 16%, about 2% to about 14%, about 2% to about 12%, about 2% to about 10%, about 2% to about 8%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, about 3% to about 95%, about 3% to about 90%, about 3% to about 85%, about 3% to about 80%, about 3% to about 75%, about 3% to about 70%, about 3% to about 65%, about 3% to about 60%, about 3% to about 55%, about 3% to about 50%, about 3% to about 45%, about 3% to about 40%, about 3% to about 35%, about 3% to about 30%, about 3% to about 25%, about 3% to about 20%, about 3% to about 18%, about 3% to about 16%, about 3% to about 14%, about 3% to about 12%, about 3% to about 10%, about 3% to about 8%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 95%, about 4% to about 90%, about 4% to about 85%, about 4% to about 80%, about 4% to about 75%, about 4% to about 70%, about 4% to about 65%, about 4% to about 60%, about 4% to about 55%, about 4% to about 50%, about 4% to about 45%, about 4% to about 40%, about 4% to about 35%, about 4% to about 30%, about 4% to about 25%, about 4% to about 20%, about 4% to about 18%, about 4% to about 16%, about 4% to about 14%, about 4% to about 12%, about 4% to about 10%, about 4% to about 8%, about 4% to about 6%, about 4% to about 5%, about 5% to about 95%, about 5% to about 90%, about 5% to about 85%, about 5% to about 80%, about 5% to about 75%, about 5% to about 70%, about 5% to about 65%, about 5% to about 60%, about 5% to about 55%, about 5% to about 50%, about 5% to about 45%, about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 5% to about 20%, about 5% to about 18%, about 5% to about 16%, about 5% to about 14%, about 5% to about 12%, about 5% to about 10%, about 5% to about 8%, about 5% to about 6%, about 6% to about 95%, about 6% to about 90%, about 6% to about 85%, about 6% to about 80%, about 6% to about 75%, about 6% to about 70%, about 6% to about 65%, about 6% to about 60%, about 6% to about 55%, about 6% to about 50%, about 6% to about 45%, about 6% to about 40%, about 6% to about 35%, about 6% to about 30%, about 6% to about 25%, about 6% to about 20%, about 6% to about 18%, about 6% to about 16%, about 6% to about 14%, about 6% to about 12%, about 6% to about 10%, about 6% to about 8%, about 8% to about 95%, about 8% to about 90%, about 8% to about 85%, about 8% to about 80%, about 8% to about 75%, about 8% to about 70%, about 8% to about 65%, about 8% to about 60%, about 8% to about 55%, about 8% to about 50%, about 8% to about 45%, about 8% to about 40%, about 8% to about 35%, about 8% to about 30%, about 8% to about 25%, about 8% to about 20%, about 8% to about 18%, about 8% to about 16%, about 8% to about 14%, about 8% to about 12%, about 8% to about 10%, about 10% to about 95%, about 10% to about 90%, about 10% to about 85%, about 10% to about 80%, about 10% to about 75%, about 10% to about 70%, about 10% to about 65%, about 10% to about 60%, about 10% to about 55%, about 10% to about 50%, about 10% to about 45%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 25%, about 10% to about 20%, about 10% to about 18%, about 10% to about 16%, about 10% to about 14%, about 10% to about 12%, about 12% to about 95%, about 12% to about 90%, about 12% to about 85%, about 12% to about 80%, about 12% to about 75%, about 12% to about 70%, about 12% to about 65%, about 12% to about 60%, about 12% to about 55%, about 12% to about 50%, about 12% to about 45%, about 12% to about 40%, about 12% to about 35%, about 12% to about 30%, about 12% to about 25%, about 12% to about 20%, about 12% to about 18%, about 12% to about 16%, about 12% to about 14%, about 14% to about 95%, about 14% to about 90%, about 14% to about 85%, about 14% to about 80%, about 14% to about 75%, about 14% to about 70%, about 14% to about 65%, about 14% to about 60%, about 14% to about 55%, about 14% to about 50%, about 14% to about 45%, about 14% to about 40%, about 14% to about 35%, about 14% to about 30%, about 14% to about 25%, about 14% to about 20%, about 14% to about 18%, about 14% to about 16%, about 16% to about 95%, about 16% to about 90%, about 16% to about 85%, about 16% to about 80%, about 16% to about 75%, about 16% to about 70%, about 16% to about 65%, about 16% to about 60%, about 16% to about 55%, about 16% to about 50%, about 16% to about 45%, about 16% to about 40%, about 16% to about 35%, about 16% to about 30%, about 16% to about 25%, about 16% to about 20%, about 16% to about 18%, about 18% to about 95%, about 18% to about 90%, about 18% to about 85%, about 18% to about 80%, about 18% to about 75%, about 18% to about 70%, about 18% to about 65%, about 18% to about 60%, about 18% to about 55%, about 18% to about 50%, about 18% to about 45%, about 18% to about 40%, about 18% to about 35%, about 18% to about 30%, about 18% to about 25%, about 18% to about 20%, about 20% to about 95%, about 20% to about 90%, about 20% to about 85%, about 20% to about 80%, about 20% to about 75%, about 20% to about 70%, about 20% to about 65%, about 20% to about 60%, about 20% to about 55%, about 20% to about 50%, about 20% to about 45%, about 20% to about 40%, about 20% to about 35%, about 20% to about 30%, about 20% to about 25%, about 25% to about 95%, about 25% to about 90%, about 25% to about 85%, about 25% to about 80%, about 25% to about 75%, about 25% to about 70%, about 25% to about 65%, about 25% to about 60%, about 25% to about 55%, about 25% to about 50%, about 25% to about 45%, about 25% to about 40%, about 25% to about 35%, about 25% to about 30%, about 30% to about 95%, about 30% to about 90%, about 30% to about 85%, about 30% to about 80%, about 30% to about 75%, about 30% to about 70%, about 30% to about 65%, about 30% to about 60%, about 30% to about 55%, about 30% to about 50%, about 30% to about 45%, about 30% to about 40%, about 30% to about 35%, about 35% to about 95%, about 35% to about 90%, about 35% to about 85%, about 35% to about 80%, about 35% to about 75%, about 35% to about 70%, about 35% to about 65%, about 35% to about 60%, about 35% to about 55%, about 35% to about 50%, about 35% to about 45%, about 35% to about 40%, about 40% to about 95%, about 40% to about 90%, about 40% to about 85%, about 40% to about 80%, about 40% to about 75%, about 40% to about 70%, about 40% to about 65%, about 40% to about 60%, about 40% to about 55%, about 40% to about 50%, about 40% to about 45%, about 45% to about 95%, about 45% to about 90%, about 45% to about 85%, about 45% to about 80%, about 45% to about 75%, about 45% to about 70%, about 45% to about 65%, about 45% to about 60%, about 45% to about 55%, about 45% to about 50%, about 50% to about 95%, about 50% to about 90%, about 50% to about 85%, about 50% to about 80%, about 50% to about 75%, about 50% to about 70%, about 50% to about 65%, about 50% to about 60%, about 50% to about 55%, about 55% to about 95%, about 55% to about 90%, about 55% to about 85%, about 55% to about 80%, about 55% to about 75%, about 55% to about 70%, about 55% to about 65%, about 55% to about 60%, about 60% to about 95%, about 60% to about 90%, about 60% to about 85%, about 60% to about 80%, about 60% to about 75%, about 60% to about 70%, about 60% to about 65%, about 65% to about 95%, about 65% to about 90%, about 65% to about 85%, about 65% to about 80%, about 65% to about 75%, about 65% to about 70%, about 70% to about 95%, about 70% to about 90%, about 70% to about 85%, about 70% to about 80%, about 70% to about 75%, about 75% to about 95%, about 75% to about 90%, about 75% to about 85%, about 75% to about 80%, about 80% to about 95%, about 80% to about 90%, about 80% to about 85%, about 85% to about 95%, about 85% to about 90%, or about 90% to about 95%.

In some embodiments of any of the pods described herein, where the bottom end comprises at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, the percentage of the total surface area of the bottom end that is represented by at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod is about 1% to about 100%, about 1% to about 95%, about 1% to about 90%, about 1% to about 85%, about 1% to about 80%, about 1% to about 75%, about 1% to about 70%, about 1% to about 65%, about 1% to about 60%, about 1% to about 55%, about 1% to about 50%, about 1% to about 45%, about 1% to about 40%, about 1% to about 35%, about 1% to about 30%, about 1% to about 25%, about 1% to about 20%, about 1% to about 18%, about 1% to about 16%, about 1% to about 14%, about 1% to about 12%, about 1% to about 10%, about 1% to about 8%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 100%, about 2% to about 95%, about 2% to about 90%, about 2% to about 85%, about 2% to about 80%, about 2% to about 75%, about 2% to about 70%, about 2% to about 65%, about 2% to about 60%, about 2% to about 55%, about 2% to about 50%, about 2% to about 45%, about 2% to about 40%, about 2% to about 35%, about 2% to about 30%, about 2% to about 25%, about 2% to about 20%, about 2% to about 18%, about 2% to about 16%, about 2% to about 14%, about 2% to about 12%, about 2% to about 10%, about 2% to about 8%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, about 3% to about 100%, about 3% to about 95%, about 3% to about 90%, about 3% to about 85%, about 3% to about 80%, about 3% to about 75%, about 3% to about 70%, about 3% to about 65%, about 3% to about 60%, about 3% to about 55%, about 3% to about 50%, about 3% to about 45%, about 3% to about 40%, about 3% to about 35%, about 3% to about 30%, about 3% to about 25%, about 3% to about 20%, about 3% to about 18%, about 3% to about 16%, about 3% to about 14%, about 3% to about 12%, about 3% to about 10%, about 3% to about 8%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 100%, about 4% to about 95%, about 4% to about 90%, about 4% to about 85%, about 4% to about 80%, about 4% to about 75%, about 4% to about 70%, about 4% to about 65%, about 4% to about 60%, about 4% to about 55%, about 4% to about 50%, about 4% to about 45%, about 4% to about 40%, about 4% to about 35%, about 4% to about 30%, about 4% to about 25%, about 4% to about 20%, about 4% to about 18%, about 4% to about 16%, about 4% to about 14%, about 4% to about 12%, about 4% to about 10%, about 4% to about 8%, about 4% to about 6%, about 4% to about 5%, about 5% to about 100%, about 5% to about 95%, about 5% to about 90%, about 5% to about 85%, about 5% to about 80%, about 5% to about 75%, about 5% to about 70%, about 5% to about 65%, about 5% to about 60%, about 5% to about 55%, about 5% to about 50%, about 5% to about 45%, about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 5% to about 20%, about 5% to about 18%, about 5% to about 16%, about 5% to about 14%, about 5% to about 12%, about 5% to about 10%, about 5% to about 8%, about 5% to about 6%, about 6% to about 100%, about 6% to about 95%, about 6% to about 90%, about 6% to about 85%, about 6% to about 80%, about 6% to about 75%, about 6% to about 70%, about 6% to about 65%, about 6% to about 60%, about 6% to about 55%, about 6% to about 50%, about 6% to about 45%, about 6% to about 40%, about 6% to about 35%, about 6% to about 30%, about 6% to about 25%, about 6% to about 20%, about 6% to about 18%, about 6% to about 16%, about 6% to about 14%, about 6% to about 12%, about 6% to about 10%, about 6% to about 8%, about 8% to about 100%, about 8% to about 95%, about 8% to about 90%, about 8% to about 85%, about 8% to about 80%, about 8% to about 75%, about 8% to about 70%, about 8% to about 65%, about 8% to about 60%, about 8% to about 55%, about 8% to about 50%, about 8% to about 45%, about 8% to about 40%, about 8% to about 35%, about 8% to about 30%, about 8% to about 25%, about 8% to about 20%, about 8% to about 18%, about 8% to about 16%, about 8% to about 14%, about 8% to about 12%, about 8% to about 10%, about 10% to about 100%, about 10% to about 95%, about 10% to about 90%, about 10% to about 85%, about 10% to about 80%, about 10% to about 75%, about 10% to about 70%, about 10% to about 65%, about 10% to about 60%, about 10% to about 55%, about 10% to about 50%, about 10% to about 45%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 25%, about 10% to about 20%, about 10% to about 18%, about 10% to about 16%, about 10% to about 14%, about 10% to about 12%, about 12% to about 100%, about 12% to about 95%, about 12% to about 90%, about 12% to about 85%, about 12% to about 80%, about 12% to about 75%, about 12% to about 70%, about 12% to about 65%, about 12% to about 60%, about 12% to about 55%, about 12% to about 50%, about 12% to about 45%, about 12% to about 40%, about 12% to about 35%, about 12% to about 30%, about 12% to about 25%, about 12% to about 20%, about 12% to about 18%, about 12% to about 16%, about 12% to about 14%, about 14% to about 100%, about 14% to about 95%, about 14% to about 90%, about 14% to about 85%, about 14% to about 80%, about 14% to about 75%, about 14% to about 70%, about 14% to about 65%, about 14% to about 60%, about 14% to about 55%, about 14% to about 50%, about 14% to about 45%, about 14% to about 40%, about 14% to about 35%, about 14% to about 30%, about 14% to about 25%, about 14% to about 20%, about 14% to about 18%, about 14% to about 16%, about 16% to about 100%, about 16% to about 95%, about 16% to about 90%, about 16% to about 85%, about 16% to about 80%, about 16% to about 75%, about 16% to about 70%, about 16% to about 65%, about 16% to about 60%, about 16% to about 55%, about 16% to about 50%, about 16% to about 45%, about 16% to about 40%, about 16% to about 35%, about 16% to about 30%, about 16% to about 25%, about 16% to about 20%, about 16% to about 18%, about 18% to about 100%, about 18% to about 95%, about 18% to about 90%, about 18% to about 85%, about 18% to about 80%, about 18% to about 75%, about 18% to about 70%, about 18% to about 65%, about 18% to about 60%, about 18% to about 55%, about 18% to about 50%, about 18% to about 45%, about 18% to about 40%, about 18% to about 35%, about 18% to about 30%, about 18% to about 25%, about 18% to about 20%, about 20% to about 100%, about 20% to about 95%, about 20% to about 90%, about 20% to about 85%, about 20% to about 80%, about 20% to about 75%, about 20% to about 70%, about 20% to about 65%, about 20% to about 60%, about 20% to about 55%, about 20% to about 50%, about 20% to about 45%, about 20% to about 40%, about 20% to about 35%, about 20% to about 30%, about 20% to about 25%, about 25% to about 100%, about 25% to about 95%, about 25% to about 90%, about 25% to about 85%, about 25% to about 80%, about 25% to about 75%, about 25% to about 70%, about 25% to about 65%, about 25% to about 60%, about 25% to about 55%, about 25% to about 50%, about 25% to about 45%, about 25% to about 40%, about 25% to about 35%, about 25% to about 30%, about 30% to about 100%, about 30% to about 95%, about 30% to about 90%, about 30% to about 85%, about 30% to about 80%, about 30% to about 75%, about 30% to about 70%, about 30% to about 65%, about 30% to about 60%, about 30% to about 55%, about 30% to about 50%, about 30% to about 45%, about 30% to about 40%, about 30% to about 35%, about 35% to about 100%, about 35% to about 95%, about 35% to about 90%, about 35% to about 85%, about 35% to about 80%, about 35% to about 75%, about 35% to about 70%, about 35% to about 65%, about 35% to about 60%, about 35% to about 55%, about 35% to about 50%, about 35% to about 45%, about 35% to about 40%, about 40% to about 100%, about 40% to about 95%, about 40% to about 90%, about 40% to about 85%, about 40% to about 80%, about 40% to about 75%, about 40% to about 70%, about 40% to about 65%, about 40% to about 60%, about 40% to about 55%, about 40% to about 50%, about 40% to about 45%, about 45% to about 100%, about 45% to about 95%, about 45% to about 90%, about 45% to about 85%, about 45% to about 80%, about 45% to about 75%, about 45% to about 70%, about 45% to about 65%, about 45% to about 60%, about 45% to about 55%, about 45% to about 50%, about 50% to about 100%, about 50% to about 95%, about 50% to about 90%, about 50% to about 85%, about 50% to about 80%, about 50% to about 75%, about 50% to about 70%, about 50% to about 65%, about 50% to about 60%, about 50% to about 55%, about 55% to about 100%, about 55% to about 95%, about 55% to about 90%, about 55% to about 85%, about 55% to about 80%, about 55% to about 75%, about 55% to about 70%, about 55% to about 65%, about 55% to about 60%, about 60% to about 100%, about 60% to about 95%, about 60% to about 90%, about 60% to about 85%, about 60% to about 80%, about 60% to about 75%, about 60% to about 70%, about 60% to about 65%, about 65% to about 100%, about 65% to about 95%, about 65% to about 90%, about 65% to about 85%, about 65% to about 80%, about 65% to about 75%, about 65% to about 70%; about 70% to about 100%, about 70% to about 95%, about 70% to about 90%, about 70% to about 85%, about 70% to about 80%, about 70% to about 75%, about 75% to about 100%, about 75% to about 95%, about 75% to about 90%, about 75% to about 85%, about 75% to about 80%, about 80% to about 100%, about 80% to about 95%, about 80% to about 90%, about 80% to about 85%, about 85% to about 100%, about 85% to about 95%, about 85% to about 90%, about 90% to about 100%, about 90% to about 95%, or about 95% to about 100%.

In some embodiments of any of the pods described herein, where the top end comprises at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, the percentage of the total surface area of the top end that is represented by at least one area covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod is about 1% to about 100%, about 1% to about 95%, about 1% to about 90%, about 1% to about 85%, about 1% to about 80%, about 1% to about 75%, about 1% to about 70%, about 1% to about 65%, about 1% to about 60%, about 1% to about 55%, about 1% to about 50%, about 1% to about 45%, about 1% to about 40%, about 1% to about 35%, about 1% to about 30%, about 1% to about 25%, about 1% to about 20%, about 1% to about 18%, about 1% to about 16%, about 1% to about 14%, about 1% to about 12%, about 1% to about 10%, about 1% to about 8%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 100%, about 2% to about 95%, about 2% to about 90%, about 2% to about 85%, about 2% to about 80%, about 2% to about 75%, about 2% to about 70%, about 2% to about 65%, about 2% to about 60%, about 2% to about 55%, about 2% to about 50%, about 2% to about 45%, about 2% to about 40%, about 2% to about 35%, about 2% to about 30%, about 2% to about 25%, about 2% to about 20%, about 2% to about 18%, about 2% to about 16%, about 2% to about 14%, about 2% to about 12%, about 2% to about 10%, about 2% to about 8%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, about 3% to about 100%, about 3% to about 95%, about 3% to about 90%, about 3% to about 85%, about 3% to about 80%, about 3% to about 75%, about 3% to about 70%, about 3% to about 65%, about 3% to about 60%, about 3% to about 55%, about 3% to about 50%, about 3% to about 45%, about 3% to about 40%, about 3% to about 35%, about 3% to about 30%, about 3% to about 25%, about 3% to about 20%, about 3% to about 18%, about 3% to about 16%, about 3% to about 14%, about 3% to about 12%, about 3% to about 10%, about 3% to about 8%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 100%, about 4% to about 95%, about 4% to about 90%, about 4% to about 85%, about 4% to about 80%, about 4% to about 75%, about 4% to about 70%, about 4% to about 65%, about 4% to about 60%, about 4% to about 55%, about 4% to about 50%, about 4% to about 45%, about 4% to about 40%, about 4% to about 35%, about 4% to about 30%, about 4% to about 25%, about 4% to about 20%, about 4% to about 18%, about 4% to about 16%, about 4% to about 14%, about 4% to about 12%, about 4% to about 10%, about 4% to about 8%, about 4% to about 6%, about 4% to about 5%, about 5% to about 100%, about 5% to about 95%, about 5% to about 90%, about 5% to about 85%, about 5% to about 80%, about 5% to about 75%, about 5% to about 70%, about 5% to about 65%, about 5% to about 60%, about 5% to about 55%, about 5% to about 50%, about 5% to about 45%, about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 5% to about 20%, about 5% to about 18%, about 5% to about 16%, about 5% to about 14%, about 5% to about 12%, about 5% to about 10%, about 5% to about 8%, about 5% to about 6%, about 6% to about 100%, about 6% to about 95%, about 6% to about 90%, about 6% to about 85%, about 6% to about 80%, about 6% to about 75%, about 6% to about 70%, about 6% to about 65%, about 6% to about 60%, about 6% to about 55%, about 6% to about 50%, about 6% to about 45%, about 6% to about 40%, about 6% to about 35%, about 6% to about 30%, about 6% to about 25%, about 6% to about 20%, about 6% to about 18%, about 6% to about 16%, about 6% to about 14%, about 6% to about 12%, about 6% to about 10%, about 6% to about 8%, about 8% to about 100%, about 8% to about 95%, about 8% to about 90%, about 8% to about 85%, about 8% to about 80%, about 8% to about 75%, about 8% to about 70%, about 8% to about 65%, about 8% to about 60%, about 8% to about 55%, about 8% to about 50%, about 8% to about 45%, about 8% to about 40%, about 8% to about 35%, about 8% to about 30%, about 8% to about 25%, about 8% to about 20%, about 8% to about 18%, about 8% to about 16%, about 8% to about 14%, about 8% to about 12%, about 8% to about 10%, about 10% to about 100%, about 10% to about 95%, about 10% to about 90%, about 10% to about 85%, about 10% to about 80%, about 10% to about 75%, about 10% to about 70%, about 10% to about 65%, about 10% to about 60%, about 10% to about 55%, about 10% to about 50%, about 10% to about 45%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 25%, about 10% to about 20%, about 10% to about 18%, about 10% to about 16%, about 10% to about 14%, about 10% to about 12%, about 12% to about 100%, about 12% to about 95%, about 12% to about 90%, about 12% to about 85%, about 12% to about 80%, about 12% to about 75%, about 12% to about 70%, about 12% to about 65%, about 12% to about 60%, about 12% to about 55%, about 12% to about 50%, about 12% to about 45%, about 12% to about 40%, about 12% to about 35%, about 12% to about 30%, about 12% to about 25%, about 12% to about 20%, about 12% to about 18%, about 12% to about 16%, about 12% to about 14%, about 14% to about 100%, about 14% to about 95%, about 14% to about 90%, about 14% to about 85%, about 14% to about 80%, about 14% to about 75%, about 14% to about 70%, about 14% to about 65%, about 14% to about 60%, about 14% to about 55%, about 14% to about 50%, about 14% to about 45%, about 14% to about 40%, about 14% to about 35%, about 14% to about 30%, about 14% to about 25%, about 14% to about 20%, about 14% to about 18%, about 14% to about 16%, about 16% to about 100%, about 16% to about 95%, about 16% to about 90%, about 16% to about 85%, about 16% to about 80%, about 16% to about 75%, about 16% to about 70%, about 16% to about 65%, about 16% to about 60%, about 16% to about 55%, about 16% to about 50%, about 16% to about 45%, about 16% to about 40%, about 16% to about 35%, about 16% to about 30%, about 16% to about 25%, about 16% to about 20%, about 16% to about 18%, about 18% to about 100%, about 18% to about 95%, about 18% to about 90%, about 18% to about 85%, about 18% to about 80%, about 18% to about 75%, about 18% to about 70%, about 18% to about 65%, about 18% to about 60%, about 18% to about 55%, about 18% to about 50%, about 18% to about 45%, about 18% to about 40%, about 18% to about 35%, about 18% to about 30%, about 18% to about 25%, about 18% to about 20%, about 20% to about 100%, about 20% to about 95%, about 20% to about 90%, about 20% to about 85%, about 20% to about 80%, about 20% to about 75%, about 20% to about 70%, about 20% to about 65%, about 20% to about 60%, about 20% to about 55%, about 20% to about 50%, about 20% to about 45%, about 20% to about 40%, about 20% to about 35%, about 20% to about 30%, about 20% to about 25%, about 25% to about 100%, about 25% to about 95%, about 25% to about 90%, about 25% to about 85%, about 25% to about 80%, about 25% to about 75%, about 25% to about 70%, about 25% to about 65%, about 25% to about 60%, about 25% to about 55%, about 25% to about 50%, about 25% to about 45%, about 25% to about 40%, about 25% to about 35%, about 25% to about 30%, about 30% to about 100%, about 30% to about 95%, about 30% to about 90%, about 30% to about 85%, about 30% to about 80%, about 30% to about 75%, about 30% to about 70%, about 30% to about 65%, about 30% to about 60%, about 30% to about 55%, about 30% to about 50%, about 30% to about 45%, about 30% to about 40%, about 30% to about 35%, about 35% to about 100%, about 35% to about 95%, about 35% to about 90%, about 35% to about 85%, about 35% to about 80%, about 35% to about 75%, about 35% to about 70%, about 35% to about 65%, about 35% to about 60%, about 35% to about 55%, about 35% to about 50%, about 35% to about 45%, about 35% to about 40%, about 40% to about 100%, about 40% to about 95%, about 40% to about 90%, about 40% to about 85%, about 40% to about 80%, about 40% to about 75%, about 40% to about 70%, about 40% to about 65%, about 40% to about 60%, about 40% to about 55%, about 40% to about 50%, about 40% to about 45%, about 45% to about 100%, about 45% to about 95%, about 45% to about 90%, about 45% to about 85%, about 45% to about 80%, about 45% to about 75%, about 45% to about 70%, about 45% to about 65%, about 45% to about 60%, about 45% to about 55%, about 45% to about 50%, about 50% to about 100%, about 50% to about 95%, about 50% to about 90%, about 50% to about 85%, about 50% to about 80%, about 50% to about 75%, about 50% to about 70%, about 50% to about 65%, about 50% to about 60%, about 50% to about 55%, about 55% to about 100%, about 55% to about 95%, about 55% to about 90%, about 55% to about 85%, about 55% to about 80%, about 55% to about 75%, about 55% to about 70%, about 55% to about 65%, about 55% to about 60%, about 60% to about 100%, about 60% to about 95%, about 60% to about 90%, about 60% to about 85%, about 60% to about 80%, about 60% to about 75%, about 60% to about 70%, about 60% to about 65%, about 65% to about 100%, about 65% to about 95%, about 65% to about 90%, about 65% to about 85%, about 65% to about 80%, about 65% to about 75%, about 65% to about 70%; about 70% to about 100%, about 70% to about 95%, about 70% to about 90%, about 70% to about 85%, about 70% to about 80%, about 70% to about 75%, about 75% to about 100%, about 75% to about 95%, about 75% to about 90%, about 75% to about 85%, about 75% to about 80%, about 80% to about 100%, about 80% to about 95%, about 80% to about 90%, about 80% to about 85%, about 85% to about 100%, about 85% to about 95%, about 85% to about 90%, about 90% to about 100%, about 90% to about 95%, or about 95% to about 100%. Containers

In some examples, the bottom end of the container is flat, has a hemispherical shape, or has a conical shape.

In some examples of any of the pods described herein, the container has an interior volume of about 0.05 mL to about 1,000 mL, about 0.05 mL to about 950 mL, about 0.05 mL to about 900 mL, about 0.05 mL to about 850 mL, about 0.05 mL to about 800 mL, about 0.05 mL to about 750 mL, about 0.05 mL to about 700 mL, about 0.05 mL to about 650 mL, about 0.05 mL to about 600 mL, about 0.05 mL to about 550 mL, about 0.05 mL to about 500 mL, about 0.05 mL to about 450 mL, about 0.05 mL to about 400 mL, about 0.05 mL to about 350 mL, about 0.05 mL to about 300 mL, about 0.05 mL to about 250 mL, about 0.05 mL to about 200 mL, about 0.05 mL to about 150 mL, about 0.05 mL to about 100 mL, about 0.05 mL to about 90 mL, about 0.05 mL to about 80 mL, about 0.05 mL to about 70 mL, about 0.05 mL to about 60 mL, about 0.05 mL to about 50 mL, about 0.05 mL to about 45 mL, about 0.05 mL to about 40 mL, about 0.05 mL to about 35 mL, about 0.05 mL to about 30 mL, about 0.05 mL to about 25 mL, about 0.05 mL to about 20 mL, about 0.05 mL to about 15 mL, about 0.05 mL to about 10 mL, about 0.05 mL to about 8 mL, about 0.05 mL to about 6 mL, about 0.05 mL to about 5 mL, about 0.05 mL to about 4 mL, about 0.05 mL to about 3 mL, about 0.05 mL to about 2 mL, about 0.05 mL to about 1 mL, about 0.05 mL to about 0.5 mL, about 0.05 mL to about 0.1 mL, about 0.1 mL to about 1,000 mL, about 0.1 mL to about 950 mL, about 0.1 mL to about 900 mL, about 0.1 mL to about 850 mL, about 0.1 mL to about 800 mL, about 0.1 mL to about 750 mL, about 0.1 mL to about 700 mL, about 0.1 mL to about 650 mL, about 0.1 mL to about 600 mL, about 0.1 mL to about 550 mL, about 0.1 mL to about 500 mL, about 0.1 mL to about 450 mL, about 0.1 mL to about 400 mL, about 0.1 mL to about 350 mL, about 0.1 mL to about 300 mL, about 0.1 mL to about 250 mL, about 0.1 mL to about 200 mL, about 0.1 mL to about 150 mL, about 0.1 mL to about 100 mL, about 0.1 mL to about 90 mL, about 0.1 mL to about 80 mL, about 0.1 mL to about 70 mL, about 0.1 mL to about 60 mL, about 0.1 mL to about 50 mL, about 0.1 mL to about 45 mL, about 0.1 mL to about 40 mL, about 0.1 mL to about 35 mL, about 0.1 mL to about 30 mL, about 0.1 mL to about 25 mL, about 0.1 mL to about 20 mL, about 0.1 mL to about 15 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 8 mL, about 0.1 mL to about 6 mL, about 0.1 mL to about 5 mL, about 0.1 mL to about 4 mL, about 0.1 mL to about 3 mL, about 0.1 mL to about 2 mL, about 0.1 mL to about 1 mL, about 0.1 mL to about 0.5 mL, about 0.5 mL to about 1,000 mL, about 0.5 mL to about 950 mL, about 0.5 mL to about 900 mL, about 0.5 mL to about 850 mL, about 0.5 mL to about 800 mL, about 0.5 mL to about 750 mL, about 0.5 mL to about 700 mL, about 0.5 mL to about 650 mL, about 0.5 mL to about 600 mL, about 0.5 mL to about 550 mL, about 0.5 mL to about 500 mL, about 0.5 mL to about 450 mL, about 0.5 mL to about 400 mL, about 0.5 mL to about 350 mL, about 0.5 mL to about 300 mL, about 0.5 mL to about 250 mL, about 0.5 mL to about 200 mL, about 0.5 mL to about 150 mL, about 0.5 mL to about 100 mL, about 0.5 mL to about 90 mL, about 0.5 mL to about 80 mL, about 0.5 mL to about 70 mL, about 0.5 mL to about 60 mL, about 0.5 mL to about 50 mL, about 0.5 mL to about 45 mL, about 0.5 mL to about 40 mL, about 0.5 mL to about 35 mL, about 0.5 mL to about 30 mL, about 0.5 mL to about 25 mL, about 0.5 mL to about 20 mL, about 0.5 mL to about 15 mL, about 0.5 mL to about 10 mL, about 0.5 mL to about 8 mL, about 0.5 mL to about 6 mL, about 0.5 mL to about 5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1 mL, about 1 mL to about 1,000 mL, about 1 mL to about 950 mL, about 1 mL to about 900 mL, about 1 mL to about 850 mL, about 1 mL to about 800 mL, about 1 mL to about 750 mL, about 1 mL to about 700 mL, about 1 mL to about 650 mL, about 1 mL to about 600 mL, about 1 mL to about 550 mL, about 1 mL to about 500 mL, about 1 mL to about 450 mL, about 1 mL to about 400 mL, about 1 mL to about 350 mL, about 1 mL to about 300 mL, about 1 mL to about 250 mL, about 1 mL to about 200 mL, about 1 mL to about 150 mL, about 1 mL to about 100 mL, about 1 mL to about 90 mL, about 1 mL to about 80 mL, about 1 mL to about 70 mL, about 1 mL to about 60 mL, about 1 mL to about 50 mL, about 1 mL to about 45 mL, about 1 mL to about 40 mL, about 1 mL to about 35 mL, about 1 mL to about 30 mL, about 1 mL to about 25 mL, about 1 mL to about 20 mL, about 1 mL to about 15 mL, about 1 mL to about 10 mL, about 1 mL to about 8 mL, about 1 mL to about 6 mL, about 1 mL to about 5 mL, about 1 mL to about 4 mL, about 1 mL to about 3 mL, about 1 mL to about 2 mL, about 2 mL to about 1,000 mL, about 2 mL to about 950 mL, about 2 mL to about 900 mL, about 2 mL to about 850 mL, about 2 mL to about 800 mL, about 2 mL to about 750 mL, about 2 mL to about 700 mL, about 2 mL to about 650 mL, about 2 mL to about 600 mL, about 2 mL to about 550 mL, about 2 mL to about 500 mL, about 2 mL to about 450 mL, about 2 mL to about 400 mL, about 2 mL to about 350 mL, about 2 mL to about 300 mL, about 2 mL to about 250 mL, about 2 mL to about 200 mL, about 2 mL to about 150 mL, about 2 mL to about 100 mL, about 2 mL to about 90 mL, about 2 mL to about 80 mL, about 2 mL to about 70 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 45 mL, about 2 mL to about 40 mL, about 2 mL to about 35 mL, about 2 mL to about 30 mL, about 2 mL to about 25 mL, about 2 mL to about 20 mL, about 2 mL to about 15 mL, about 2 mL to about 10 mL, about 2 mL to about 8 mL, about 2 mL to about 6 mL, about 2 mL to about 5 mL, about 2 mL to about 4 mL, about 2 mL to about 3 mL, about 3 mL to about 1,000 mL, about 3 mL to about 950 mL, about 3 mL to about 900 mL, about 3 mL to about 850 mL, about 3 mL to about 800 mL, about 3 mL to about 750 mL, about 3 mL to about 700 mL, about 3 mL to about 650 mL, about 3 mL to about 600 mL, about 3 mL to about 550 mL, about 3 mL to about 500 mL, about 3 mL to about 450 mL, about 3 mL to about 400 mL, about 3 mL to about 350 mL, about 3 mL to about 300 mL, about 3 mL to about 250 mL, about 3 mL to about 200 mL, about 3 mL to about 150 mL, about 3 mL to about 100 mL, about 3 mL to about 90 mL, about 3 mL to about 80 mL, about 3 mL to about 70 mL, about 3 mL to about 60 mL, about 3 mL to about 50 mL, about 3 mL to about 45 mL, about 3 mL to about 40 mL, about 3 mL to about 35 mL, about 3 mL to about 30 mL, about 3 mL to about 25 mL, about 3 mL to about 20 mL, about 3 mL to about 15 mL, about 3 mL to about 10 mL, about 3 mL to about 8 mL, about 3 mL to about 6 mL, about 3 mL to about 5 mL, about 3 mL to about 4 mL, about 4 mL to about 1,000 mL, about 4 mL to about 950 mL, about 4 mL to about 900 mL, about 4 mL to about 850 mL, about 4 mL to about 800 mL, about 4 mL to about 750 mL, about 4 mL to about 700 mL, about 4 mL to about 650 mL, about 4 mL to about 600 mL, about 4 mL to about 550 mL, about 4 mL to about 500 mL, about 4 mL to about 450 mL, about 4 mL to about 400 mL, about 4 mL to about 350 mL, about 4 mL to about 300 mL, about 4 mL to about 250 mL, about 4 mL to about 200 mL, about 4 mL to about 150 mL, about 4 mL to about 100 mL, about 4 mL to about 90 mL, about 4 mL to about 80 mL, about 4 mL to about 70 mL, about 4 mL to about 60 mL, about 4 mL to about 50 mL, about 4 mL to about 45 mL, about 4 mL to about 40 mL, about 4 mL to about 35 mL, about 4 mL to about 30 mL, about 4 mL to about 25 mL, about 4 mL to about 20 mL, about 4 mL to about 15 mL, about 4 mL to about 10 mL, about 4 mL to about 8 mL, about 4 mL to about 6 mL, about 4 mL to about 5 mL, about 5 mL to about 1,000 mL, about 5 mL to about 950 mL, about 5 mL to about 900 mL, about 5 mL to about 850 mL, about 5 mL to about 800 mL, about 5 mL to about 750 mL, about 5 mL to about 700 mL, about 5 mL to about 650 mL, about 5 mL to about 600 mL, about 5 mL to about 550 mL, about 5 mL to about 500 mL, about 5 mL to about 450 mL, about 5 mL to about 400 mL, about 5 mL to about 350 mL, about 5 mL to about 300 mL, about 5 mL to about 250 mL, about 5 mL to about 200 mL, about 5 mL to about 150 mL, about 5 mL to about 100 mL, about 5 mL to about 90 mL, about 5 mL to about 80 mL, about 5 mL to about 70 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 45 mL, about 5 mL to about 40 mL, about 5 mL to about 35 mL, about 5 mL to about 30 mL, about 5 mL to about 25 mL, about 5 mL to about 20 mL, about 5 mL to about 15 mL, about 5 mL to about 10 mL, about 5 mL to about 8 mL, about 5 mL to about 6 mL, about 6 mL to about 1,000 mL, about 6 mL to about 950 mL, about 6 mL to about 900 mL, about 6 mL to about 850 mL, about 6 mL to about 800 mL, about 6 mL to about 750 mL, about 6 mL to about 700 mL, about 6 mL to about 650 mL, about 6 mL to about 600 mL, about 6 mL to about 550 mL, about 6 mL to about 500 mL, about 6 mL to about 450 mL, about 6 mL to about 400 mL, about 6 mL to about 350 mL, about 6 mL to about 300 mL, about 6 mL to about 250 mL, about 6 mL to about 200 mL, about 6 mL to about 150 mL, about 6 mL to about 100 mL, about 6 mL to about 90 mL, about 6 mL to about 80 mL, about 6 mL to about 70 mL, about 6 mL to about 60 mL, about 6 mL to about 50 mL, about 6 mL to about 45 mL, about 6 mL to about 40 mL, about 6 mL to about 35 mL, about 6 mL to about 30 mL, about 6 mL to about 25 mL, about 6 mL to about 20 mL, about 6 mL to about 15 mL, about 6 mL to about 10 mL, about 6 mL to about 8 mL, about 8 mL to about 1,000 mL, about 8 mL to about 950 mL, about 8 mL to about 900 mL, about 8 mL to about 850 mL, about 8 mL to about 800 mL, about 8 mL to about 750 mL, about 8 mL to about 700 mL, about 8 mL to about 650 mL, about 8 mL to about 600 mL, about 8 mL to about 550 mL, about 8 mL to about 500 mL, about 8 mL to about 450 mL, about 8 mL to about 400 mL, about 8 mL to about 350 mL, about 8 mL to about 300 mL, about 8 mL to about 250 mL, about 8 mL to about 200 mL, about 8 mL to about 150 mL, about 8 mL to about 100 mL, about 8 mL to about 90 mL, about 8 mL to about 80 mL, about 8 mL to about 70 mL, about 8 mL to about 60 mL, about 8 mL to about 50 mL, about 8 mL to about 45 mL, about 8 mL to about 40 mL, about 8 mL to about 35 mL, about 8 mL to about 30 mL, about 8 mL to about 25 mL, about 8 mL to about 20 mL, about 8 mL to about 15 mL, about 8 mL to about 10 mL, about 10 mL to about 1,000 mL, about 10 mL to about 950 mL, about 10 mL to about 900 mL, about 10 mL to about 850 mL, about 10 mL to about 800 mL, about 10 mL to about 750 mL, about 10 mL to about 700 mL, about 10 mL to about 650 mL, about 10 mL to about 600 mL, about 10 mL to about 550 mL, about 10 mL to about 500 mL, about 10 mL to about 450 mL, about 10 mL to about 400 mL, about 10 mL to about 350 mL, about 10 mL to about 300 mL, about 10 mL to about 250 mL, about 10 mL to about 200 mL, about 10 mL to about 150 mL, about 10 mL to about 100 mL, about 10 mL to about 90 mL, about 10 mL to about 80 mL, about 10 mL to about 70 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 45 mL, about 10 mL to about 40 mL, about 10 mL to about 35 mL, about 10 mL to about 30 mL, about 10 mL to about 25 mL, about 10 mL to about 20 mL, about 10 mL to about 15 mL, about 15 mL to about 1,000 mL, about 15 mL to about 950 mL, about 15 mL to about 900 mL, about 15 mL to about 850 mL, about 15 mL to about 800 mL, about 15 mL to about 750 mL, about 15 mL to about 700 mL, about 15 mL to about 650 mL, about 15 mL to about 600 mL, about 15 mL to about 550 mL, about 15 mL to about 500 mL, about 15 mL to about 450 mL, about 15 mL to about 400 mL, about 15 mL to about 350 mL, about 15 mL to about 300 mL, about 15 mL to about 250 mL, about 15 mL to about 200 mL, about 15 mL to about 150 mL, about 15 mL to about 100 mL, about 15 mL to about 90 mL, about 15 mL to about 80 mL, about 15 mL to about 70 mL, about 15 mL to about 60 mL, about 15 mL to about 50 mL, about 15 mL to about 45 mL, about 15 mL to about 40 mL, about 15 mL to about 35 mL, about 15 mL to about 30 mL, about 15 mL to about 25 mL, about 15 mL to about 20 mL, about 20 mL to about 1,000 mL, about 20 mL to about 950 mL, about 20 mL to about 900 mL, about 20 mL to about 850 mL, about 20 mL to about 800 mL, about 20 mL to about 750 mL, about 20 mL to about 700 mL, about 20 mL to about 650 mL, about 20 mL to about 600 mL, about 20 mL to about 550 mL, about 20 mL to about 500 mL, about 20 mL to about 450 mL, about 20 mL to about 400 mL, about 20 mL to about 350 mL, about 20 mL to about 300 mL, about 20 mL to about 250 mL, about 20 mL to about 200 mL, about 20 mL to about 150 mL, about 20 mL to about 100 mL, about 20 mL to about 90 mL, about 20 mL to about 80 mL, about 20 mL to about 70 mL, about 20 mL to about 60 mL, about 20 mL to about 50 mL, about 20 mL to about 45 mL, about 20 mL to about 40 mL, about 20 mL to about 35 mL, about 20 mL to about 30 mL, about 20 mL to about 25 mL, about 25 mL to about 1,000 mL, about 25 mL to about 950 mL, about 25 mL to about 900 mL, about 25 mL to about 850 mL, about 25 mL to about 800 mL, about 25 mL to about 750 mL, about 25 mL to about 700 mL, about 25 mL to about 650 mL, about 25 mL to about 600 mL, about 25 mL to about 550 mL, about 25 mL to about 500 mL, about 25 mL to about 450 mL, about 25 mL to about 400 mL, about 25 mL to about 350 mL, about 25 mL to about 300 mL, about 25 mL to about 250 mL, about 25 mL to about 200 mL, about 25 mL to about 150 mL, about 25 mL to about 100 mL, about 25 mL to about 90 mL, about 25 mL to about 80 mL, about 25 mL to about 70 mL, about 25 mL to about 60 mL, about 25 mL to about 50 mL, about 25 mL to about 45 mL, about 25 mL to about 40 mL, about 25 mL to about 35 mL, about 25 mL to about 30 mL, about 30 mL to about 1,000 mL, about 30 mL to about 950 mL, about 30 mL to about 900 mL, about 30 mL to about 850 mL, about 30 mL to about 800 mL, about 30 mL to about 750 mL, about 30 mL to about 700 mL, about 30 mL to about 650 mL, about 30 mL to about 600 mL, about 30 mL to about 550 mL, about 30 mL to about 500 mL, about 30 mL to about 450 mL, about 30 mL to about 400 mL, about 30 mL to about 350 mL, about 30 mL to about 300 mL, about 30 mL to about 250 mL, about 30 mL to about 200 mL, about 30 mL to about 150 mL, about 30 mL to about 100 mL, about 30 mL to about 90 mL, about 30 mL to about 80 mL, about 30 mL to about 70 mL, about 30 mL to about 60 mL, about 30 mL to about 50 mL, about 30 mL to about 45 mL, about 30 mL to about 40 mL, about 30 mL to about 35 mL, about 35 mL to about 1,000 mL, about 35 mL to about 950 mL, about 35 mL to about 900 mL, about 35 mL to about 850 mL, about 35 mL to about 800 mL, about 35 mL to about 750 mL, about 35 mL to about 700 mL, about 35 mL to about 650 mL, about 35 mL to about 600 mL, about 35 mL to about 550 mL, about 35 mL to about 500 mL, about 35 mL to about 450 mL, about 35 mL to about 400 mL, about 35 mL to about 350 mL, about 35 mL to about 300 mL, about 35 mL to about 250 mL, about 35 mL to about 200 mL, about 35 mL to about 150 mL, about 35 mL to about 100 mL, about 35 mL to about 90 mL, about 35 mL to about 80 mL, about 35 mL to about 70 mL, about 35 mL to about 60 mL, about 35 mL to about 50 mL, about 35 mL to about 45 mL, about 35 mL to about 40 mL, about 40 mL to about 1,000 mL, about 40 mL to about 950 mL, about 40 mL to about 900 mL, about 40 mL to about 850 mL, about 40 mL to about 800 mL, about 40 mL to about 750 mL, about 40 mL to about 700 mL, about 40 mL to about 650 mL, about 40 mL to about 600 mL, about 40 mL to about 550 mL, about 40 mL to about 500 mL, about 40 mL to about 450 mL, about 40 mL to about 400 mL, about 40 mL to about 350 mL, about 40 mL to about 300 mL, about 40 mL to about 250 mL, about 40 mL to about 200 mL, about 40 mL to about 150 mL, about 40 mL to about 100 mL, about 40 mL to about 90 mL, about 40 mL to about 80 mL, about 40 mL to about 70 mL, about 40 mL to about 60 mL, about 40 mL to about 50 mL, about 40 mL to about 45 mL, about 45 mL to about 1,000 mL, about 45 mL to about 950 mL, about 45 mL to about 900 mL, about 45 mL to about 850 mL, about 45 mL to about 800 mL, about 45 mL to about 750 mL, about 45 mL to about 700 mL, about 45 mL to about 650 mL, about 45 mL to about 600 mL, about 45 mL to about 550 mL, about 45 mL to about 500 mL, about 45 mL to about 450 mL, about 45 mL to about 400 mL, about 45 mL to about 350 mL, about 45 mL to about 300 mL, about 45 mL to about 250 mL, about 45 mL to about 200 mL, about 45 mL to about 150 mL, about 45 mL to about 100 mL, about 45 mL to about 90 mL, about 45 mL to about 80 mL, about 45 mL to about 70 mL, about 45 mL to about 60 mL, about 45 mL to about 50 mL, about 50 mL to about 1,000 mL, about 50 mL to about 950 mL, about 50 mL to about 900 mL, about 50 mL to about 850 mL, about 50 mL to about 800 mL, about 50 mL to about 750 mL, about 50 mL to about 700 mL, about 50 mL to about 650 mL, about 50 mL to about 600 mL, about 50 mL to about 550 mL, about 50 mL to about 500 mL, about 50 mL to about 450 mL, about 50 mL to about 400 mL, about 50 mL to about 350 mL, about 50 mL to about 300 mL, about 50 mL to about 250 mL, about 50 mL to about 200 mL, about 50 mL to about 150 mL, about 50 mL to about 100 mL, about 50 mL to about 90 mL, about 50 mL to about 80 mL, about 50 mL to about 70 mL, about 50 mL to about 60 mL, about 60 mL to about 1,000 mL, about 60 mL to about 950 mL, about 60 mL to about 900 mL, about 60 mL to about 850 mL, about 60 mL to about 800 mL, about 60 mL to about 750 mL, about 60 mL to about 700 mL, about 60 mL to about 650 mL, about 60 mL to about 600 mL, about 60 mL to about 550 mL, about 60 mL to about 500 mL, about 60 mL to about 450 mL, about 60 mL to about 400 mL, about 60 mL to about 350 mL, about 60 mL to about 300 mL, about 60 mL to about 250 mL, about 60 mL to about 200 mL, about 60 mL to about 150 mL, about 60 mL to about 100 mL, about 60 mL to about 90 mL, about 60 mL to about 80 mL, about 60 mL to about 70 mL, about 70 mL to about 1,000 mL, about 70 mL to about 950 mL, about 70 mL to about 900 mL, about 70 mL to about 850 mL, about 70 mL to about 800 mL, about 70 mL to about 750 mL, about 70 mL to about 700 mL, about 70 mL to about 650 mL, about 70 mL to about 600 mL, about 70 mL to about 550 mL, about 70 mL to about 500 mL, about 70 mL to about 450 mL, about 70 mL to about 400 mL, about 70 mL to about 350 mL, about 70 mL to about 300 mL, about 70 mL to about 250 mL, about 70 mL to about 200 mL, about 70 mL to about 150 mL, about 70 mL to about 100 mL, about 70 mL to about 90 mL, about 70 mL to about 80 mL, about 80 mL to about 1,000 mL, about 80 mL to about 950 mL, about 80 mL to about 900 mL, about 80 mL to about 850 mL, about 80 mL to about 800 mL, about 80 mL to about 750 mL, about 80 mL to about 700 mL, about 80 mL to about 650 mL, about 80 mL to about 600 mL, about 80 mL to about 550 mL, about 80 mL to about 500 mL, about 80 mL to about 450 mL, about 80 mL to about 400 mL, about 80 mL to about 350 mL, about 80 mL to about 300 mL, about 80 mL to about 250 mL, about 80 mL to about 200 mL, about 80 mL to about 150 mL, about 80 mL to about 100 mL, about 80 mL to about 90 mL, about 90 mL to about 1,000 mL, about 90 mL to about 950 mL, about 90 mL to about 900 mL, about 90 mL to about 850 mL, about 90 mL to about 800 mL, about 90 mL to about 750 mL, about 90 mL to about 700 mL, about 90 mL to about 650 mL, about 90 mL to about 600 mL, about 90 mL to about 550 mL, about 90 mL to about 500 mL, about 90 mL to about 450 mL, about 90 mL to about 400 mL, about 90 mL to about 350 mL, about 90 mL to about 300 mL, about 90 mL to about 250 mL, about 90 mL to about 200 mL, about 90 mL to about 150 mL, about 90 mL to about 100 mL, about 100 mL to about 1,000 mL, about 100 mL to about 950 mL, about 100 mL to about 900 mL, about 100 mL to about 850 mL, about 100 mL to about 800 mL, about 100 mL to about 750 mL, about 100 mL to about 700 mL, about 100 mL to about 650 mL, about 100 mL to about 600 mL, about 100 mL to about 550 mL, about 100 mL to about 500 mL, about 100 mL to about 450 mL, about 100 mL to about 400 mL, about 100 mL to about 350 mL, about 100 mL to about 300 mL, about 100 mL to about 250 mL, about 100 mL to about 200 mL, about 100 mL to about 150 mL, about 150 mL to about 1,000 mL, about 150 mL to about 950 mL, about 150 mL to about 900 mL, about 150 mL to about 850 mL, about 150 mL to about 800 mL, about 150 mL to about 750 mL, about 150 mL to about 700 mL, about 150 mL to about 650 mL, about 150 mL to about 600 mL, about 150 mL to about 550 mL, about 150 mL to about 500 mL, about 150 mL to about 450 mL, about 150 mL to about 400 mL, about 150 mL to about 350 mL, about 150 mL to about 300 mL, about 150 mL to about 250 mL, about 150 mL to about 200 mL, about 200 mL to about 1,000 mL, about 200 mL to about 950 mL, about 200 mL to about 900 mL, about 200 mL to about 850 mL, about 200 mL to about 800 mL, about 200 mL to about 750 mL, about 200 mL to about 700 mL, about 200 mL to about 650 mL, about 200 mL to about 600 mL, about 200 mL to about 550 mL, about 200 mL to about 500 mL, about 200 mL to about 450 mL, about 200 mL to about 400 mL, about 200 mL to about 350 mL, about 200 mL to about 300 mL, about 200 mL to about 250 mL, about 250 mL to about 1,000 mL, about 250 mL to about 950 mL, about 250 mL to about 900 mL, about 250 mL to about 850 mL, about 250 mL to about 800 mL, about 250 mL to about 750 mL, about 250 mL to about 700 mL, about 250 mL to about 650 mL, about 250 mL to about 600 mL, about 250 mL to about 550 mL, about 250 mL to about 500 mL, about 250 mL to about 450 mL, about 250 mL to about 400 mL, about 250 mL to about 350 mL, about 250 mL to about 300 mL, about 300 mL to about 1,000 mL, about 300 mL to about 950 mL, about 300 mL to about 900 mL, about 300 mL to about 850 mL, about 300 mL to about 800 mL, about 300 mL to about 750 mL, about 300 mL to about 700 mL, about 300 mL to about 650 mL, about 300 mL to about 600 mL, about 300 mL to about 550 mL, about 300 mL to about 500 mL, about 300 mL to about 450 mL, about 300 mL to about 400 mL, about 300 mL to about 350 mL, about 350 mL to about 1,000 mL, about 350 mL to about 950 mL, about 350 mL to about 900 mL, about 350 mL to about 850 mL, about 350 mL to about 800 mL, about 350 mL to about 750 mL, about 350 mL to about 700 mL, about 350 mL to about 650 mL, about 350 mL to about 600 mL, about 350 mL to about 550 mL, about 350 mL to about 500 mL, about 350 mL to about 450 mL, about 350 mL to about 400 mL, about 400 mL to about 1,000 mL, about 400 mL to about 950 mL, about 400 mL to about 900 mL, about 400 mL to about 850 mL, about 400 mL to about 800 mL, about 400 mL to about 750 mL, about 400 mL to about 700 mL, about 400 mL to about 650 mL, about 400 mL to about 600 mL, about 400 mL to about 550 mL, about 400 mL to about 500 mL, about 400 mL to about 450 mL, about 450 mL to about 1,000 mL, about 450 mL to about 950 mL, about 450 mL to about 900 mL, about 450 mL to about 850 mL, about 450 mL to about 800 mL, about 450 mL to about 750 mL, about 450 mL to about 700 mL, about 450 mL to about 650 mL, about 450 mL to about 600 mL, about 450 mL to about 550 mL, about 450 mL to about 500 mL, about 500 mL to about 1,000 mL, about 500 mL to about 950 mL, about 500 mL to about 900 mL, about 500 mL to about 850 mL, about 500 mL to about 800 mL, about 500 mL to about 750 mL, about 500 mL to about 700 mL, about 500 mL to about 650 mL, about 500 mL to about 600 mL, about 500 mL to about 550 mL, about 550 mL to about 1,000 mL, about 550 mL to about 950 mL, about 550 mL to about 900 mL, about 550 mL to about 850 mL, about 550 mL to about 800 mL, about 550 mL to about 750 mL, about 550 mL to about 700 mL, about 550 mL to about 650 mL, about 550 mL to about 600 mL, about 600 mL to about 1,000 mL, about 600 mL to about 950 mL, about 600 mL to about 900 mL, about 600 mL to about 850 mL, about 600 mL to about 800 mL, about 600 mL to about 750 mL, about 600 mL to about 700 mL, about 600 mL to about 650 mL, about 650 mL to about 1,000 mL, about 650 mL to about 950 mL, about 650 mL to about 900 mL, about 650 mL to about 850 mL, about 650 mL to about 800 mL, about 650 mL to about 750 mL, about 650 mL to about 700 mL, about 700 mL to about 1,000 mL, about 700 mL to about 950 mL, about 700 mL to about 900 mL, about 700 mL to about 850 mL, about 700 mL to about 800 mL, about 700 mL to about 750 mL, about 750 mL to about 1,000 mL, about 750 mL to about 950 mL, about 750 mL to about 900 mL, about 750 mL to about 850 mL, about 750 mL to about 800 mL, about 800 mL to about 1,000 mL, about 800 mL to about 950 mL, about 800 mL to about 900 mL, about 800 mL to about 850 mL, about 850 mL to about 1,000 mL, about 850 mL to about 950 mL, about 850 mL to about 900 mL, about 900 mL to about 1,000 mL, about 900 mL to about 950 mL, or about 950 mL to about 1,000 mL. Wall

The wall can be composed or composed in part of polyethylene (PE), polypropylene (PP), polystyrene (PS), polyvinyl chloride (PVC), an acrylic polymer, a polyester, silicone rubber, silicon, a polyurethane, a halogenated plastic (e.g., chlorinated polyethylene, chlorinated polyvinyl chloride, chlorosulfonated polyethylene, polychloroprene, fluorinated ethylene propylene), biaxially-oriented polypropylene (BOPP), a bioplastic, cellophane, an ethane-derived plastic, nylon, poly-lactic acid (PLA), polyethylene terephthalate (PE or PETE plastic code 1), polypropylene, polystyrene, polycarbonate, polyeurothene, or poly(methyl methacrylate). Additional exemplary compositions that the wall can be composed of include: Bakelite (phenol-formaldehyde resin), Kevlar or Twaron (para-aramid), Kynar (polyvinylidene difluoride), Mylar (polyethylene terephthalate film), Neoprene (polychloroprene), nylon (polyamide 6,6), Orlon (polyacrylonitrile), Rilsan

(polyamide 11 & 12), Technora (copolyamide), Teflon (polytetrafluoroethylene), Ultem (polyimide), Vectran (aromatic polyamide), Viton (poly-tetrafluoroethylene), or Zylon (poly-p-phenylene-2,6-benzobisoxazole (PBO). In some embodiments of the pods, the wall can be composed or composed in part of nylon, polypropylene, or polyvinyl alcohol (PVA).

In some embodiments of the pods, the wall has a cylindrical shape, a conical shape, a rectangular shape, a square shape, a spherical shape, a hemispherical shape, an hour glass shape, or an ellipsoidal shape. Chromatography Resins

In some embodiments of any of the methods described herein, the pod further includes a chromatography resin disposed in the interior of the container. In some examples, the chromatography resin is a commercial chromatography resin. In other examples, the chromatography resin is not a commercial chromatography resin.

The chromatography resin can be any chromatography resin known in the art. For example, the chromatography resin can be an affinity chromatography resin (e.g., a protein A resin, a protein G resin, a protein L resin, a nickel resin, or a magnetic bead resin), a hydrophobic interaction chromatography resin, a mixed mode chromatography resin, a molecular sieve chromatography resin. In some embodiments, the resin is a specific affinity resin, e.g., a resin conjugated to an agent (e.g., an antibody, antibody fragment, peptide, nucleic acid, and the like) that is specific for a recombinant protein of interest. In some embodiments, the chromatography resin is an alkali-resistant resin. In some embodiments, the chromatography resin is a sterilized chromatography resin.

The chromatography resin disposed in the interior of the container can have a volume of about 0.1 mL to about 500 mL, about 0.1 mL to about 490 mL, about 0.1 mL to about 480 mL, about 0.1 mL to about 460 mL, about 0.1 mL to about 440 mL, about 0.1 mL to about 420 mL, about 0.1 mL to about 400 mL, about 0.1 mL to about 380 mL, about 0.1 mL to about 360 mL, about 0.1 mL to about 340 mL, about 0.1 mL to about 320 mL, about 0.1 mL to about 300 mL, about 0.1 mL to about 280 mL, about 0.1 mL to about 260 mL, about 0.1 mL to about 250 mL, about 0.1 mL to about 240 mL, about 0.1 mL to about 220 mL, about 0.1 mL to about 210 mL, about 0.1 mL to about 200 mL, about 0.1 mL to about 190 mL, about 0.1 mL to about 180 mL, about 0.1 mL to about 170 mL, about 0.1 mL to about 160 mL, about 0.1 mL to about 150 mL, about 0.1 mL to about 140 mL, about 0.1 mL to about 130 mL, about 0.1 mL to about 120 mL, about 0.1 mL to about 110 mL, about 0.1 mL to about 100 mL, about 0.1 mL to about 90 mL, about 0.1 mL to about 80 mL, about 0.1 mL to about 70 mL, about 0.1 mL to about 60 mL, about 0.1 mL to about 50 mL, about 0.1 mL to about 40 mL, about 0.1 mL to about 30 mL, about 0.1 mL to about 25 mL, about 0.1 mL to about 20 mL, about 0.1 mL to about 15 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 5 mL, about 0.1 mL to about 2 mL, about 0.1 mL to about 1 mL, about 0.2 mL to about 500 mL, about 0.2 mL to about 490 mL, about 0.2 mL to about 480 mL, about 0.2 mL to about 460 mL, about 0.2 mL to about 440 mL, about 0.2 mL to about 420 mL, about 0.2 mL to about 400 mL, about 0.2 mL to about 380 mL, about 0.2 mL to about 360 mL, about 0.2 mL to about 340 mL, about 0.2 mL to about 320 mL, about 0.2 mL to about 300 mL, about 0.2 mL to about 280 mL, about 0.2 mL to about 260 mL, about 0.2 mL to about 250 mL, about 0.2 mL to about 240 mL, about 0.2 mL to about 220 mL, about 0.2 mL to about 210 mL, about 0.2 mL to about 200 mL, about 0.2 mL to about 190 mL, about 0.2 mL to about 180 mL, about 0.2 mL to about 170 mL, about 0.2 mL to about 160 mL, about 0.2 mL to about 150 mL, about 0.2 mL to about 140 mL, about 0.2 mL to about 130 mL, about 0.2 mL to about 120 mL, about 0.2 mL to about 110 mL, about 0.2 mL to about 100 mL, about 0.2 mL to about 90 mL, about 0.2 mL to about 80 mL, about 0.2 mL to about 70 mL, about 0.2 mL to about 60 mL, about 0.2 mL to about 50 mL, about 0.2 mL to about 40 mL, about 0.2 mL to about 30 mL, about 0.2 mL to about 25 mL, about 0.2 mL to about 20 mL, about 0.2 mL to about 15 mL, about 0.2 mL to about 10 mL, about 0.2 mL to about 5 mL, about 0.2 mL to about 2 mL, about 0.2 mL to about 1 mL, about 0.4 mL to about 500 mL, about 0.4 mL to about 490 mL, about 0.4 mL to about 480 mL, about 0.4 mL to about 460 mL, about 0.4 mL to about 440 mL, about 0.4 mL to about 420 mL, about 0.4 mL to about 400 mL, about 0.4 mL to about 380 mL, about 0.4 mL to about 360 mL, about 0.4 mL to about 340 mL, about 0.4 mL to about 320 mL, about 0.4 mL to about 300 mL, about 0.4 mL to about 280 mL, about 0.4 mL to about 260 mL, about 0.4 mL to about 250 mL, about 0.4 mL to about 240 mL, about 0.4 mL to about 220 mL, about 0.4 mL to about 210 mL, about 0.4 mL to about 200 mL, about 0.4 mL to about 190 mL, about 0.4 mL to about 180 mL, about 0.4 mL to about 170 mL, about 0.4 mL to about 160 mL, about 0.4 mL to about 150 mL, about 0.4 mL to about 140 mL, about 0.4 mL to about 130 mL, about 0.4 mL to about 120 mL, about 0.4 mL to about 110 mL, about 0.4 mL to about 100 mL, about 0.4 mL to about 90 mL, about 0.4 mL to about 80 mL, about 0.4 mL to about 70 mL, about 0.4 mL to about 60 mL, about 0.4 mL to about 50 mL, about 0.4 mL to about 40 mL, about 0.4 mL to about 30 mL, about 0.4 mL to about 25 mL, about 0.4 mL to about 20 mL, about 0.4 mL to about 15 mL, about 0.4 mL to about 10 mL, about 0.4 mL to about 5 mL, about 0.4 mL to about 2 mL, about 0.4 mL to about 1 mL, about 0.5 mL to about 500 mL, about 0.5 mL to about 490 mL, about 0.5 mL to about 480 mL, about 0.5 mL to about 460 mL, about 0.5 mL to about 440 mL, about 0.5 mL to about 420 mL, about 0.5 mL to about 400 mL, about 0.5 mL to about 380 mL, about 0.5 mL to about 360 mL, about 0.5 mL to about 340 mL, about 0.5 mL to about 320 mL, about 0.5 mL to about 300 mL, about 0.5 mL to about 280 mL, about 0.5 mL to about 260 mL, about 0.5 mL to about 250 mL, about 0.5 mL to about 240 mL, about 0.5 mL to about 220 mL, about 0.5 mL to about 210 mL, about 0.5 mL to about 200 mL, about 0.5 mL to about 190 mL, about 0.5 mL to about 180 mL, about 0.5 mL to about 170 mL, about 0.5 mL to about 160 mL, about 0.5 mL to about 150 mL, about 0.5 mL to about 140 mL, about 0.5 mL to about 130 mL, about 0.5 mL to about 120 mL, about 0.5 mL to about 110 mL, about 0.5 mL to about 100 mL, about 0.5 mL to about 90 mL, about 0.5 mL to about 80 mL, about 0.5 mL to about 70 mL, about 0.5 mL to about 60 mL, about 0.5 mL to about 50 mL, about 0.5 mL to about 40 mL, about 0.5 mL to about 30 mL, about 0.5 mL to about 25 mL, about 0.5 mL to about 20 mL, about 0.5 mL to about 15 mL, about 0.5 mL to about 10 mL, about 0.5 mL to about 5 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1 mL, about 0.6 mL to about 500 mL, about 0.6 mL to about 490 mL, about 0.6 mL to about 480 mL, about 0.6 mL to about 460 mL, about 0.6 mL to about 440 mL, about 0.6 mL to about 420 mL, about 0.6 mL to about 400 mL, about 0.6 mL to about 380 mL, about 0.6 mL to about 360 mL, about 0.6 mL to about 340 mL, about 0.6 mL to about 320 mL, about 0.6 mL to about 300 mL, about 0.6 mL to about 280 mL, about 0.6 mL to about 260 mL, about 0.6 mL to about 250 mL, about 0.6 mL to about 240 mL, about 0.6 mL to about 220 mL, about 0.6 mL to about 210 mL, about 0.6 mL to about 200 mL, about 0.6 mL to about 190 mL, about 0.6 mL to about 180 mL, about 0.6 mL to about 170 mL, about 0.6 mL to about 160 mL, about 0.6 mL to about 150 mL, about 0.6 mL to about 140 mL, about 0.6 mL to about 130 mL, about 0.6 mL to about 120 mL, about 0.6 mL to about 110 mL, about 0.6 mL to about 100 mL, about 0.6 mL to about 90 mL, about 0.6 mL to about 80 mL, about 0.6 mL to about 70 mL, about 0.6 mL to about 60 mL, about 0.6 mL to about 50 mL, about 0.6 mL to about 40 mL, about 0.6 mL to about 30 mL, about 0.6 mL to about 25 mL, about 0.6 mL to about 20 mL, about 0.6 mL to about 15 mL, about 0.6 mL to about 10 mL, about 0.6 mL to about 5 mL, about 0.6 mL to about 2 mL, about 0.6 mL to about 1 mL, about 0.8 mL to about 500 mL, about 0.8 mL to about 490 mL, about 0.8 mL to about 480 mL, about 0.8 mL to about 460 mL, about 0.8 mL to about 440 mL, about 0.8 mL to about 420 mL, about 0.8 mL to about 400 mL, about 0.8 mL to about 380 mL, about 0.8 mL to about 360 mL, about 0.8 mL to about 340 mL, about 0.8 mL to about 320 mL, about 0.8 mL to about 300 mL, about 0.8 mL to about 280 mL, about 0.8 mL to about 260 mL, about 0.8 mL to about 250 mL, about 0.8 mL to about 240 mL, about 0.8 mL to about 220 mL, about 0.8 mL to about 210 mL, about 0.8 mL to about 200 mL, about 0.8 mL to about 190 mL, about 0.8 mL to about 180 mL, about 0.8 mL to about 170 mL, about 0.8 mL to about 160 mL, about 0.8 mL to about 150 mL, about 0.8 mL to about 140 mL, about 0.8 mL to about 130 mL, about 0.8 mL to about 120 mL, about 0.8 mL to about 110 mL, about 0.8 mL to about 100 mL, about 0.8 mL to about 90 mL, about 0.8 mL to about 80 mL, about 0.8 mL to about 70 mL, about 0.8 mL to about 60 mL, about 0.8 mL to about 50 mL, about 0.8 mL to about 40 mL, about 0.8 mL to about 30 mL, about 0.8 mL to about 25 mL, about 0.8 mL to about 20 mL, about 0.8 mL to about 15 mL, about 0.8 mL to about 10 mL, about 0.8 mL to about 5 mL, about 0.8 mL to about 2 mL, about 0.8 mL to about 1 mL, about 1 mL to about 500 mL, about 1 mL to about 490 mL, about 1 mL to about 480 mL, about 1 mL to about 460 mL, about 1 mL to about 440 mL, about 1 mL to about 420 mL, about 1 mL to about 400 mL, about 1 mL to about 380 mL, about 1 mL to about 360 mL, about 1 mL to about 340 mL, about 1 mL to about 320 mL, about 1 mL to about 300 mL, about 1 mL to about 280 mL, about 1 mL to about 260 mL, about 1 mL to about 250 mL, about 1 mL to about 240 mL, about 1 mL to about 220 mL, about 1 mL to about 210 mL, about 1 mL to about 200 mL, about 1 mL to about 190 mL, about 1 mL to about 180 mL, about 1 mL to about 170 mL, about 1 mL to about 160 mL, about 1 mL to about 150 mL, about 1 mL to about 140 mL, about 1 mL to about 130 mL, about 1 mL to about 120 mL, about 1 mL to about 110 mL, about 1 mL to about 100 mL, about 1 mL to about 90 mL, about 1 mL to about 80 mL, about 1 mL to about 70 mL, about 1 mL to about 60 mL, about 1 mL to about 50 mL, about 1 mL to about 40 mL, about 1 mL to about 30 mL, about 1 mL to about 25 mL, about 1 mL to about 20 mL, about 1 mL to about 15 mL, about 1 mL to about 10 mL, about 1 mL to about 5 mL, about 1 mL to about 2 mL, about 2 mL to about 500 mL, about 2 mL to about 490 mL, about 2 mL to about 480 mL, about 2 mL to about 460 mL, about 2 mL to about 440 mL, about 2 mL to about 420 mL, about 2 mL to about 400 mL, about 2 mL to about 380 mL, about 2 mL to about 360 mL, about 2 mL to about 340 mL, about 2 mL to about 320 mL, about 2 mL to about 300 mL, about 2 mL to about 280 mL, about 2 mL to about 260 mL, about 2 mL to about 250 mL, about 2 mL to about 240 mL, about 2 mL to about 220 mL, about 2 mL to about 210 mL, about 2 mL to about 200 mL, about 2 mL to about 190 mL, about 2 mL to about 180 mL, about 2 mL to about 170 mL, about 2 mL to about 160 mL, about 2 mL to about 150 mL, about 2 mL to about 140 mL, about 2 mL to about 130 mL, about 2 mL to about 120 mL, about 2 mL to about 110 mL, about 2 mL to about 100 mL, about 2 mL to about 90 mL, about 2 mL to about 80 mL, about 2 mL to about 70 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 40 mL, about 2 mL to about 30 mL, about 2 mL to about 25 mL, about 2 mL to about 20 mL, about 2 mL to about 15 mL, about 2 mL to about 10 mL, about 2 mL to about 5 mL, about 5 mL to about 500 mL, about 5 mL to about 490 mL, about 5 mL to about 480 mL, about 5 mL to about 460 mL, about 5 mL to about 440 mL, about 5 mL to about 420 mL, about 5 mL to about 400 mL, about 5 mL to about 380 mL, about 5 mL to about 360 mL, about 5 mL to about 340 mL, about 5 mL to about 320 mL, about 5 mL to about 300 mL, about 5 mL to about 280 mL, about 5 mL to about 260 mL, about 5 mL to about 250 mL, about 5 mL to about 240 mL, about 5 mL to about 220 mL, about 5 mL to about 210 mL, about 5 mL to about 200 mL, about 5 mL to about 190 mL, about 5 mL to about 180 mL, about 5 mL to about 170 mL, about 5 mL to about 160 mL, about 5 mL to about 150 mL, about 5 mL to about 140 mL, about 5 mL to about 130 mL, about 5 mL to about 120 mL, about 5 mL to about 110 mL, about 5 mL to about 100 mL, about 5 mL to about 90 mL, about 5 mL to about 80 mL, about 5 mL to about 70 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 40 mL, about 5 mL to about 30 mL, about 5 mL to about 25 mL, about 5 mL to about 20 mL, about 5 mL to about 15 mL, about 5 mL to about 10 mL, about 10 mL to about 500 mL, about 10 mL to about 490 mL, about 10 mL to about 480 mL, about 10 mL to about 460 mL, about 10 mL to about 440 mL, about 10 mL to about 420 mL, about 10 mL to about 400 mL, about 10 mL to about 380 mL, about 10 mL to about 360 mL, about 10 mL to about 340 mL, about 10 mL to about 320 mL, about 10 mL to about 300 mL, about 10 mL to about 280 mL, about 10 mL to about 260 mL, about 10 mL to about 250 mL, about 10 mL to about 240 mL, about 10 mL to about 220 mL, about 10 mL to about 210 mL, about 10 mL to about 200 mL, about 10 mL to about 190 mL, about 10 mL to about 180 mL, about 10 mL to about 170 mL, about 10 mL to about 160 mL, about 10 mL to about 150 mL, about 10 mL to about 140 mL, about 10 mL to about 130 mL, about 10 mL to about 120 mL, about 10 mL to about 110 mL, about 10 mL to about 100 mL, about 10 mL to about 90 mL, about 10 mL to about 80 mL, about 10 mL to about 70 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 40 mL, about 10 mL to about 30 mL, about 10 mL to about 25 mL, about 10 mL to about 20 mL, about 10 mL to about 15 mL, about 15 mL to about 500 mL, about 15 mL to about 490 mL, about 15 mL to about 480 mL, about 15 mL to about 460 mL, about 15 mL to about 440 mL, about 15 mL to about 420 mL, about 15 mL to about 400 mL, about 15 mL to about 380 mL, about 15 mL to about 360 mL, about 15 mL to about 340 mL, about 15 mL to about 320 mL, about 15 mL to about 300 mL, about 15 mL to about 280 mL, about 15 mL to about 260 mL, about 15 mL to about 250 mL, about 15 mL to about 240 mL, about 15 mL to about 220 mL, about 15 mL to about 210 mL, about 15 mL to about 200 mL, about 15 mL to about 190 mL, about 15 mL to about 180 mL, about 15 mL to about 170 mL, about 15 mL to about 160 mL, about 15 mL to about 150 mL, about 15 mL to about 140 mL, about 15 mL to about 130 mL, about 15 mL to about 120 mL, about 15 mL to about 110 mL, about 15 mL to about 100 mL, about 15 mL to about 90 mL, about 15 mL to about 80 mL, about 15 mL to about 70 mL, about 15 mL to about 60 mL, about 15 mL to about 50 mL, about 15 mL to about 40 mL, about 15 mL to about 30 mL, about 15 mL to about 25 mL, about 15 mL to about 20 mL, about 20 mL to about 500 mL, about 20 mL to about 490 mL, about 20 mL to about 480 mL, about 20 mL to about 460 mL, about 20 mL to about 440 mL, about 20 mL to about 420 mL, about 20 mL to about 400 mL, about 20 mL to about 380 mL, about 20 mL to about 360 mL, about 20 mL to about 340 mL, about 20 mL to about 320 mL, about 20 mL to about 300 mL, about 20 mL to about 280 mL, about 20 mL to about 260 mL, about 20 mL to about 250 mL, about 20 mL to about 240 mL, about 20 mL to about 220 mL, about 20 mL to about 210 mL, about 20 mL to about 200 mL, about 20 mL to about 190 mL, about 20 mL to about 180 mL, about 20 mL to about 170 mL, about 20 mL to about 160 mL, about 20 mL to about 150 mL, about 20 mL to about 140 mL, about 20 mL to about 130 mL, about 20 mL to about 120 mL, about 20 mL to about 110 mL, about 20 mL to about 100 mL, about 20 mL to about 90 mL, about 20 mL to about 80 mL, about 20 mL to about 70 mL, about 20 mL to about 60 mL, about 20 mL to about 50 mL, about 20 mL to about 40 mL, about 20 mL to about 30 mL, about 20 mL to about 25 mL, about 25 mL to about 500 mL, about 25 mL to about 490 mL, about 25 mL to about 480 mL, about 25 mL to about 460 mL, about 25 mL to about 440 mL, about 25 mL to about 420 mL, about 25 mL to about 400 mL, about 25 mL to about 380 mL, about 25 mL to about 360 mL, about 25 mL to about 340 mL, about 25 mL to about 320 mL, about 25 mL to about 300 mL, about 25 mL to about 280 mL, about 25 mL to about 260 mL, about 25 mL to about 250 mL, about 25 mL to about 240 mL, about 25 mL to about 220 mL, about 25 mL to about 210 mL, about 25 mL to about 200 mL, about 25 mL to about 190 mL, about 25 mL to about 180 mL, about 25 mL to about 170 mL, about 25 mL to about 160 mL, about 25 mL to about 150 mL, about 25 mL to about 140 mL, about 25 mL to about 130 mL, about 25 mL to about 120 mL, about 25 mL to about 110 mL, about 25 mL to about 100 mL, about 25 mL to about 90 mL, about 25 mL to about 80 mL, about 25 mL to about 70 mL, about 25 mL to about 60 mL, about 25 mL to about 50 mL, about 25 mL to about 40 mL, about 25 mL to about 30 mL, about 30 mL to about 500 mL, about 30 mL to about 490 mL, about 30 mL to about 480 mL, about 30 mL to about 460 mL, about 30 mL to about 440 mL, about 30 mL to about 420 mL, about 30 mL to about 400 mL, about 30 mL to about 380 mL, about 30 mL to about 360 mL, about 30 mL to about 340 mL, about 30 mL to about 320 mL, about 30 mL to about 300 mL, about 30 mL to about 280 mL, about 30 mL to about 260 mL, about 30 mL to about 250 mL, about 30 mL to about 240 mL, about 30 mL to about 220 mL, about 30 mL to about 210 mL, about 30 mL to about 200 mL, about 30 mL to about 190 mL, about 30 mL to about 180 mL, about 30 mL to about 170 mL, about 30 mL to about 160 mL, about 30 mL to about 150 mL, about 30 mL to about 140 mL, about 30 mL to about 130 mL, about 30 mL to about 120 mL, about 30 mL to about 110 mL, about 30 mL to about 100 mL, about 30 mL to about 90 mL, about 30 mL to about 80 mL, about 30 mL to about 70 mL, about 30 mL to about 60 mL, about 30 mL to about 50 mL, about 30 mL to about 40 mL, about 40 mL to about 500 mL, about 40 mL to about 490 mL, about 40 mL to about 480 mL, about 40 mL to about 460 mL, about 40 mL to about 440 mL, about 40 mL to about 420 mL, about 40 mL to about 400 mL, about 40 mL to about 380 mL, about 40 mL to about 360 mL, about 40 mL to about 340 mL, about 40 mL to about 320 mL, about 40 mL to about 300 mL, about 40 mL to about 280 mL, about 40 mL to about 260 mL, about 40 mL to about 250 mL, about 40 mL to about 240 mL, about 40 mL to about 220 mL, about 40 mL to about 210 mL, about 40 mL to about 200 mL, about 40 mL to about 190 mL, about 40 mL to about 180 mL, about 40 mL to about 170 mL, about 40 mL to about 160 mL, about 40 mL to about 150 mL, about 40 mL to about 140 mL, about 40 mL to about 130 mL, about 40 mL to about 120 mL, about 40 mL to about 110 mL, about 40 mL to about 100 mL, about 40 mL to about 90 mL, about 40 mL to about 80 mL, about 40 mL to about 70 mL, about 40 mL to about 60 mL, about 40 mL to about 50 mL, about 50 mL to about 500 mL, about 50 mL to about 490 mL, about 50 mL to about 480 mL, about 50 mL to about 460 mL, about 50 mL to about 440 mL, about 50 mL to about 420 mL, about 50 mL to about 400 mL, about 50 mL to about 380 mL, about 50 mL to about 360 mL, about 50 mL to about 340 mL, about 50 mL to about 320 mL, about 50 mL to about 300 mL, about 50 mL to about 280 mL, about 50 mL to about 260 mL, about 50 mL to about 250 mL, about 50 mL to about 240 mL, about 50 mL to about 220 mL, about 50 mL to about 210 mL, about 50 mL to about 200 mL, about 50 mL to about 190 mL, about 50 mL to about 180 mL, about 50 mL to about 170 mL, about 50 mL to about 160 mL, about 50 mL to about 150 mL, about 50 mL to about 140 mL, about 50 mL to about 130 mL, about 50 mL to about 120 mL, about 50 mL to about 110 mL, about 50 mL to about 100 mL, about 50 mL to about 90 mL, about 50 mL to about 80 mL, about 50 mL to about 70 mL, about 50 mL to about 60 mL, about 60 mL to about 500 mL, about 60 mL to about 490 mL, about 60 mL to about 480 mL, about 60 mL to about 460 mL, about 60 mL to about 440 mL, about 60 mL to about 420 mL, about 60 mL to about 400 mL, about 60 mL to about 380 mL, about 60 mL to about 360 mL, about 60 mL to about 340 mL, about 60 mL to about 320 mL, about 60 mL to about 300 mL, about 60 mL to about 280 mL, about 60 mL to about 260 mL, about 60 mL to about 250 mL, about 60 mL to about 240 mL, about 60 mL to about 220 mL, about 60 mL to about 210 mL, about 60 mL to about 200 mL, about 60 mL to about 190 mL, about 60 mL to about 180 mL, about 60 mL to about 170 mL, about 60 mL to about 160 mL, about 60 mL to about 150 mL, about 60 mL to about 140 mL, about 60 mL to about 130 mL, about 60 mL to about 120 mL, about 60 mL to about 110 mL, about 60 mL to about 100 mL, about 60 mL to about 90 mL, about 60 mL to about 80 mL, about 60 mL to about 70 mL, about 70 mL to about 500 mL, about 70 mL to about 490 mL, about 70 mL to about 480 mL, about 70 mL to about 460 mL, about 70 mL to about 440 mL, about 70 mL to about 420 mL, about 70 mL to about 400 mL, about 70 mL to about 380 mL, about 70 mL to about 360 mL, about 70 mL to about 340 mL, about 70 mL to about 320 mL, about 70 mL to about 300 mL, about 70 mL to about 280 mL, about 70 mL to about 260 mL, about 70 mL to about 250 mL, about 70 mL to about 240 mL, about 70 mL to about 220 mL, about 70 mL to about 210 mL, about 70 mL to about 200 mL, about 70 mL to about 190 mL, about 70 mL to about 180 mL, about 70 mL to about 170 mL, about 70 mL to about 160 mL, about 70 mL to about 150 mL, about 70 mL to about 140 mL, about 70 mL to about 130 mL, about 70 mL to about 120 mL, about 70 mL to about 110 mL, about 70 mL to about 100 mL, about 70 mL to about 90 mL, about 70 mL to about 80 mL, about 80 mL to about 500 mL, about 80 mL to about 490 mL, about 80 mL to about 480 mL, about 80 mL to about 460 mL, about 80 mL to about 440 mL, about 80 mL to about 420 mL, about 80 mL to about 400 mL, about 80 mL to about 380 mL, about 80 mL to about 360 mL, about 80 mL to about 340 mL, about 80 mL to about 320 mL, about 80 mL to about 300 mL, about 80 mL to about 280 mL, about 80 mL to about 260 mL, about 80 mL to about 250 mL, about 80 mL to about 240 mL, about 80 mL to about 220 mL, about 80 mL to about 210 mL, about 80 mL to about 200 mL, about 80 mL to about 190 mL, about 80 mL to about 180 mL, about 80 mL to about 170 mL, about 80 mL to about 160 mL, about 80 mL to about 150 mL, about 80 mL to about 140 mL, about 80 mL to about 130 mL, about 80 mL to about 120 mL, about 80 mL to about 110 mL, about 80 mL to about 100 mL, about 80 mL to about 90 mL, about 90 mL to about 500 mL, about 90 mL to about 490 mL, about 90 mL to about 480 mL, about 90 mL to about 460 mL, about 90 mL to about 440 mL, about 90 mL to about 420 mL, about 90 mL to about 400 mL, about 90 mL to about 380 mL, about 90 mL to about 360 mL, about 90 mL to about 340 mL, about 90 mL to about 320 mL, about 90 mL to about 300 mL, about 90 mL to about 280 mL, about 90 mL to about 260 mL, about 90 mL to about 250 mL, about 90 mL to about 240 mL, about 90 mL to about 220 mL, about 90 mL to about 210 mL, about 90 mL to about 200 mL, about 90 mL to about 190 mL, about 90 mL to about 180 mL, about 90 mL to about 170 mL, about 90 mL to about 160 mL, about 90 mL to about 150 mL, about 90 mL to about 140 mL, about 90 mL to about 130 mL, about 90 mL to about 120 mL, about 90 mL to about 110 mL, about 90 mL to about 100 mL, about 100 mL to about 500 mL, about 100 mL to about 490 mL, about 100 mL to about 480 mL, about 100 mL to about 460 mL, about 100 mL to about 440 mL, about 100 mL to about 420 mL, about 100 mL to about 400 mL, about 100 mL to about 380 mL, about 100 mL to about 360 mL, about 100 mL to about 340 mL, about 100 mL to about 320 mL, about 100 mL to about 300 mL, about 100 mL to about 280 mL, about 100 mL to about 260 mL, about 100 mL to about 250 mL, about 100 mL to about 240 mL, about 100 mL to about 220 mL, about 100 mL to about 210 mL, about 100 mL to about 200 mL, about 100 mL to about 190 mL, about 100 mL to about 180 mL, about 100 mL to about 170 mL, about 100 mL to about 160 mL, about 100 mL to about 150 mL, about 100 mL to about 140 mL, about 100 mL to about 130 mL, about 100 mL to about 120 mL, about 100 mL to about 110 mL, about 110 mL to about 500 mL, about 110 mL to about 490 mL, about 110 mL to about 480 mL, about 110 mL to about 460 mL, about 110 mL to about 440 mL, about 110 mL to about 420 mL, about 110 mL to about 400 mL, about 110 mL to about 380 mL, about 110 mL to about 360 mL, about 110 mL to about 340 mL, about 110 mL to about 320 mL, about 110 mL to about 300 mL, about 110 mL to about 280 mL, about 110 mL to about 260 mL, about 110 mL to about 250 mL, about 110 mL to about 240 mL, about 110 mL to about 220 mL, about 110 mL to about 210 mL, about 110 mL to about 200 mL, about 110 mL to about 190 mL, about 110 mL to about 180 mL, about 110 mL to about 170 mL, about 110 mL to about 160 mL, about 110 mL to about 150 mL, about 110 mL to about 140 mL, about 110 mL to about 130 mL, about 110 mL to about 120 mL, about 120 mL to about 500 mL, about 120 mL to about 490 mL, about 120 mL to about 480 mL, about 120 mL to about 460 mL, about 120 mL to about 440 mL, about 120 mL to about 420 mL, about 120 mL to about 400 mL, about 120 mL to about 380 mL, about 120 mL to about 360 mL, about 120 mL to about 340 mL, about 120 mL to about 320 mL, about 120 mL to about 300 mL, about 120 mL to about 280 mL, about 120 mL to about 260 mL, about 120 mL to about 250 mL, about 120 mL to about 240 mL, about 120 mL to about 220 mL, about 120 mL to about 210 mL, about 120 mL to about 200 mL, about 120 mL to about 190 mL, about 120 mL to about 180 mL, about 120 mL to about 170 mL, about 120 mL to about 160 mL, about 120 mL to about 150 mL, about 120 mL to about 140 mL, about 120 mL to about 130 mL, about 130 mL to about 500 mL, about 130 mL to about 490 mL, about 130 mL to about 480 mL, about 130 mL to about 460 mL, about 130 mL to about 440 mL, about 130 mL to about 420 mL, about 130 mL to about 400 mL, about 130 mL to about 380 mL, about 130 mL to about 360 mL, about 130 mL to about 340 mL, about 130 mL to about 320 mL, about 130 mL to about 300 mL, about 130 mL to about 280 mL, about 130 mL to about 260 mL, about 130 mL to about 250 mL, about 130 mL to about 240 mL, about 130 mL to about 220 mL, about 130 mL to about 210 mL, about 130 mL to about 200 mL, about 130 mL to about 190 mL, about 130 mL to about 180 mL, about 130 mL to about 170 mL, about 130 mL to about 160 mL, about 130 mL to about 150 mL, about 130 mL to about 140 mL, about 140 mL to about 500 mL, about 140 mL to about 490 mL, about 140 mL to about 480 mL, about 140 mL to about 460 mL, about 140 mL to about 440 mL, about 140 mL to about 420 mL, about 140 mL to about 400 mL, about 140 mL to about 380 mL, about 140 mL to about 360 mL, about 140 mL to about 340 mL, about 140 mL to about 320 mL, about 140 mL to about 300 mL, about 140 mL to about 280 mL, about 140 mL to about 260 mL, about 140 mL to about 250 mL, about 140 mL to about 240 mL, about 140 mL to about 220 mL, about 140 mL to about 210 mL, about 140 mL to about 200 mL, about 140 mL to about 190 mL, about 140 mL to about 180 mL, about 140 mL to about 170 mL, about 140 mL to about 160 mL, about 140 mL to about 150 mL, about 150 mL to about 500 mL, about 150 mL to about 490 mL, about 150 mL to about 480 mL, about 150 mL to about 460 mL, about 150 mL to about 440 mL, about 150 mL to about 420 mL, about 150 mL to about 400 mL, about 150 mL to about 380 mL, about 150 mL to about 360 mL, about 150 mL to about 340 mL, about 150 mL to about 320 mL, about 150 mL to about 300 mL, about 150 mL to about 280 mL, about 150 mL to about 260 mL, about 150 mL to about 250 mL, about 150 mL to about 240 mL, about 150 mL to about 220 mL, about 150 mL to about 210 mL, about 150 mL to about 200 mL, about 150 mL to about 190 mL, about 150 mL to about 180 mL, about 150 mL to about 170 mL, about 150 mL to about 160 mL, about 160 mL to about 500 mL, about 160 mL to about 490 mL, about 160 mL to about 480 mL, about 160 mL to about 460 mL, about 160 mL to about 440 mL, about 160 mL to about 420 mL, about 160 mL to about 400 mL, about 160 mL to about 380 mL, about 160 mL to about 360 mL, about 160 mL to about 340 mL, about 160 mL to about 320 mL, about 160 mL to about 300 mL, about 160 mL to about 280 mL, about 160 mL to about 260 mL, about 160 mL to about 250 mL, about 160 mL to about 240 mL, about 160 mL to about 220 mL, about 160 mL to about 210 mL, about 160 mL to about 200 mL, about 160 mL to about 190 mL, about 160 mL to about 180 mL, about 160 mL to about 170 mL, about 170 mL to about 500 mL, about 170 mL to about 490 mL, about 170 mL to about 480 mL, about 170 mL to about 460 mL, about 170 mL to about 440 mL, about 170 mL to about 420 mL, about 170 mL to about 400 mL, about 170 mL to about 380 mL, about 170 mL to about 360 mL, about 170 mL to about 340 mL, about 170 mL to about 320 mL, about 170 mL to about 300 mL, about 170 mL to about 280 mL, about 170 mL to about 260 mL, about 170 mL to about 250 mL, about 170 mL to about 240 mL, about 170 mL to about 220 mL, about 170 mL to about 210 mL, about 170 mL to about 200 mL, about 170 mL to about 190 mL, about 170 mL to about 180 mL, about 180 mL to about 500 mL, about 180 mL to about 490 mL, about 180 mL to about 480 mL, about 180 mL to about 460 mL, about 180 mL to about 440 mL, about 180 mL to about 420 mL, about 180 mL to about 400 mL, about 180 mL to about 380 mL, about 180 mL to about 360 mL, about 180 mL to about 340 mL, about 180 mL to about 320 mL, about 180 mL to about 300 mL, about 180 mL to about 280 mL, about 180 mL to about 260 mL, about 180 mL to about 250 mL, about 180 mL to about 240 mL, about 180 mL to about 220 mL, about 180 mL to about 210 mL, about 180 mL to about 200 mL, about 180 mL to about 190 mL, about 190 mL to about 500 mL, about 190 mL to about 490 mL, about 190 mL to about 480 mL, about 190 mL to about 460 mL, about 190 mL to about 440 mL, about 190 mL to about 420 mL, about 190 mL to about 400 mL, about 190 mL to about 380 mL, about 190 mL to about 360 mL, about 190 mL to about 340 mL, about 190 mL to about 320 mL, about 190 mL to about 300 mL, about 190 mL to about 280 mL, about 190 mL to about 260 mL, about 190 mL to about 250 mL, about 190 mL to about 240 mL, about 190 mL to about 220 mL, about 190 mL to about 210 mL, about 190 mL to about 200 mL, about 200 mL to about 500 mL, about 200 mL to about 490 mL, about 200 mL to about 480 mL, about 200 mL to about 460 mL, about 200 mL to about 440 mL, about 200 mL to about 420 mL, about 200 mL to about 400 mL, about 200 mL to about 380 mL, about 200 mL to about 360 mL, about 200 mL to about 340 mL, about 200 mL to about 320 mL, about 200 mL to about 300 mL, about 200 mL to about 280 mL, about 200 mL to about 260 mL, about 200 mL to about 250 mL, about 200 mL to about 240 mL, about 200 mL to about 220 mL, about 200 mL to about 210 mL, about 210 mL to about 500 mL, about 210 mL to about 490 mL, about 210 mL to about 480 mL, about 210 mL to about 460 mL, about 210 mL to about 440 mL, about 210 mL to about 420 mL, about 210 mL to about 400 mL, about 210 mL to about 380 mL, about 210 mL to about 360 mL, about 210 mL to about 340 mL, about 210 mL to about 320 mL, about 210 mL to about 300 mL, about 210 mL to about 280 mL, about 210 mL to about 260 mL, about 210 mL to about 250 mL, about 210 mL to about 240 mL, about 210 mL to about 220 mL, about 220 mL to about 500 mL, about 220 mL to about 490 mL, about 220 mL to about 480 mL, about 220 mL to about 460 mL, about 220 mL to about 440 mL, about 220 mL to about 420 mL, about 220 mL to about 400 mL, about 220 mL to about 380 mL, about 220 mL to about 360 mL, about 220 mL to about 340 mL, about 220 mL to about 320 mL, about 220 mL to about 300 mL, about 220 mL to about 280 mL, about 220 mL to about 260 mL, about 220 mL to about 250 mL, about 220 mL to about 240 mL, about 240 mL to about 500 mL, about 240 mL to about 490 mL, about 240 mL to about 480 mL, about 240 mL to about 460 mL, about 240 mL to about 440 mL, about 240 mL to about 420 mL, about 240 mL to about 400 mL, about 240 mL to about 380 mL, about 240 mL to about 360 mL, about 240 mL to about 340 mL, about 240 mL to about 320 mL, about 240 mL to about 300 mL, about 240 mL to about 280 mL, about 240 mL to about 260 mL, about 240 mL to about 250 mL, about 250 mL to about 500 mL, about 250 mL to about 490 mL, about 250 mL to about 480 mL, about 250 mL to about 460 mL, about 250 mL to about 440 mL, about 250 mL to about 420 mL, about 250 mL to about 400 mL, about 250 mL to about 380 mL, about 250 mL to about 360 mL, about 250 mL to about 340 mL, about 250 mL to about 320 mL, about 250 mL to about 300 mL, about 250 mL to about 280 mL, about 250 mL to about 260 mL, about 260 mL to about 500 mL, about 260 mL to about 490 mL, about 260 mL to about 480 mL, about 260 mL to about 460 mL, about 260 mL to about 440 mL, about 260 mL to about 420 mL, about 260 mL to about 400 mL, about 260 mL to about 380 mL, about 260 mL to about 360 mL, about 260 mL to about 340 mL, about 260 mL to about 320 mL, about 260 mL to about 300 mL, about 260 mL to about 280 mL, about 280 mL to about 500 mL, about 280 mL to about 490 mL, about 280 mL to about 480 mL, about 280 mL to about 460 mL, about 280 mL to about 440 mL, about 280 mL to about 420 mL, about 280 mL to about 400 mL, about 280 mL to about 380 mL, about 280 mL to about 360 mL, about 280 mL to about 340 mL, about 280 mL to about 320 mL, about 280 mL to about 300 mL, about 300 mL to about 500 mL, about 300 mL to about 490 mL, about 300 mL to about 480 mL, about 300 mL to about 460 mL, about 300 mL to about 440 mL, about 300 mL to about 420 mL, about 300 mL to about 400 mL, about 300 mL to about 380 mL, about 300 mL to about 360 mL, about 300 mL to about 340 mL, about 300 mL to about 320 mL, about 320 mL to about 500 mL, about 320 mL to about 490 mL, about 320 mL to about 480 mL, about 320 mL to about 460 mL, about 320 mL to about 440 mL, about 320 mL to about 420 mL, about 320 mL to about 400 mL, about 320 mL to about 380 mL, about 320 mL to about 360 mL, about 320 mL to about 340 mL, about 340 mL to about 500 mL, about 340 mL to about 490 mL, about 340 mL to about 480 mL, about 340 mL to about 460 mL, about 340 mL to about 440 mL, about 340 mL to about 420 mL, about 340 mL to about 400 mL, about 340 mL to about 380 mL, about 340 mL to about 360 mL, about 360 mL to about 500 mL, about 360 mL to about 490 mL, about 360 mL to about 480 mL, about 360 mL to about 460 mL, about 360 mL to about 440 mL, about 360 mL to about 420 mL, about 360 mL to about 400 mL, about 360 mL to about 380 mL, about 380 mL to about 500 mL, about 380 mL to about 490 mL, about 380 mL to about 480 mL, about 380 mL to about 460 mL, about 380 mL to about 440 mL, about 380 mL to about 420 mL, about 380 mL to about 400 mL, about 400 mL to about 500 mL, about 400 mL to about 490 mL, about 400 mL to about 480 mL, about 400 mL to about 460 mL, about 400 mL to about 440 mL, about 400 mL to about 420 mL, about 420 mL to about 500 mL, about 420 mL to about 490 mL, about 420 mL to about 480 mL, about 420 mL to about 460 mL, about 420 mL to about 440 mL, about 440 mL to about 500 mL, about 440 mL to about 490 mL, about 440 mL to about 480 mL, about 440 mL to about 460 mL, about 460 mL to about 500 mL, about 460 mL to about 490 mL, about 460 mL to about 480 mL, about 480 mL to about 500 mL, about 480 mL to about 490 mL, or about 490 mL to about 500 mL.

In some embodiments, the chromatography resin disposed in the pod has a sterility assurance level of about 1 x 10 ^-6 to about 1 x 10 ^-10 , about 1 x 10 ^-6 to about 0.5 x 10 ^-10 , about 1 x 10 ^-6 to about 1 x 10 ^-9 , about 1 x 10 ^-6 to about 0.5 x 10 ^-9 , about 1 x 10 ^-6 to about 1 x 10 ^-8 , about 1 x 10 ^-6 to about 0.5 x 10 ^-8 , about 1 x 10 ^-6 to about 1 x 10- ⁷ , about 1 x 10 ^-6 to about 0.5 x 10 ^-7 , about 0.5 x 10 ^-7 to about 1 x 10 ^-10 , about 0.5 x 10 ^-7 to about 0.5 x 10 ^-10 , about 0.5 x 10 ^-7 to about 1 x 10 ^-9 , about 0.5 x 10 ^-7 to about 0.5 x 10 ^-9 , about 0.5 x 10 ^-7 to about 1 x 10 ^-8 , about 0.5 x 10 ^-7 to about 0.5 x 10 ^-8 , about 0.5 x 10 ^-7 to about 1 x 10 ^-7 , about 1 x 10 ^-7 to about 1 x 10 ^-10 , about 1 x 10 ^-7 to about 0.5 x 10 ^-10 , about 0.5 x 10 ^-7 to about 1 x 10 ^-9 , about 1 x 10 ^-7 to about 0.5 x 10 ^-9 , about 1 x 10 ^-7 to about 1 x 10 ^-8 , about 1 x 10 ^-7 to about 0.5 x 10 ^-8 , about 0.5 x 10 ^-8 to about 1 x 10 ^-10 , about 0.5 x 10 ^-8 to about 0.5 x 10 ^-10 , about 0.5 x 10 ^-8 to about 1 x 10 ^-9 , about 0.5 x 10 ^-8 to about 0.5 x 10 ^-9 , about 0.5 x 10 ^-8 to about 1 x 10 ^-8 , about 1 x 10 ^-8 to about 1 x 10 ^-10 , about 1 x 10 ^-8 to about 0.5 x 10 ^-10 , about 1 x 10 ^-8 to about 1 x 10 ^-9 , about 1 x 10- ⁸ to about 0.5 x 10 ^-9 , about 0.5 x 10 ^-9 to about 1 x 10 ^-10 , about 0.5 x 10 ^-9 to about 0.5 x 10 ^-10 , about 0.5 x 10 ^-9 to about 1 x 10 ^-9 , about 1 x 10 ^-9 to about 1 x 10 ^-10 , about 1 x 10- ⁹ to about 0.5 x 10 ^-10 , or about 0.5 x 10 ^-10 to about 1 x 10 ^-10 . Kits

Also provided herein are kits that include any of the pods described herein. Also provided herein are kits that include (a) any of the pods described herein (where the pod includes one or more areas covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, in the wall of the container), and (b) one or more panels that can be secured over each of the one or more areas in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the wall is prevented. Also provided herein are kits that include (a) any of the pods described herein (where the pod includes one or more areas covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, in the wall of the container), and (b) an outer sleeve that can be secured over the pod in order to cover the one or more areas in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented.

Also provided are kits that include (a) any of the pods described herein and (b) one or both of an elution outlet that can be stably fixed onto the bottom end, and a buffer reservoir that can be stably fixed onto the top end. Also provided herein are kits that include (a) any of the pods described herein (where the pod includes one or more areas covered by a semi-porous or porous material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod, in the wall of the container), and (b) a movable outer sleeve connected to the wall, such that rotation of the movable sleeve can cover the one or more areas in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented. Also provided herein are kits that include (a) any of the pods described herein and (b) a means for stably positioning the long axis of the pod in a horizontal position in the interior of a culture vessel.

Any of the kits described herein can include a means for stably positioning the pod in the interior of a culture vessel. Exemplary means for stably positioning the pod in the interior of a culture vessel (e.g., any of the culture vessels described herein) can include a bracket, a brace, and an adapter that fits over an opening in a culture vessel and allows for the insertion of the pod into the culture vessel (without dropping the pod into the culture vessel. In some embodiments, the means for stably positioning the pod in the interior of a culture vessel (e.g., any of the culture vessels described herein) is flexible and allows the pod to move with movement of the liquid culture without dropping into the culture vessel. In some embodiments, the means for stably positioning the pod in the interior of a culture vessel (e.g., any of the culture vessels described herein) is rigid and allows the pod to withstand movement of the liquid culture without dropping into the culture vessel. Non-limiting examples of means for stably positioning the pod in the interior of a culture vessel (e.g., any of the culture vessels described herein) are shown in Figures 10-19, 36, and 37. In some embodiments, the pod is stably fixed by taping the pod into the interior of a culture vessel. In some embodiments, the pod is stably fixed by snapping the pod in the interior of a culture vessel.

In some embodiments, the kit includes an adapter manifold to allow the compilation of multiple pods into a plate array, e.g., to further facilitate automated, high-throughput purification approaches.

Non-limiting examples of culture vessels include a multi-well plate (e.g., a 6- well plate, a 12-well plate, an 18-well plate, a 24-well plate, a 48-well plate, a 96-well plate, or a 384-well plate), shake flask (e.g., an Erlenmeyer flask or a T-flask), a shake tube (e.g., an Eppendorf tube), a wave bag, a bioreactor (e.g., a benchtop bioreactor, a perfusion bioreactor, a fed-batch bioreactor), or a rolling tube bioreactor. Additional examples of bioreactors are known in the art.

Some embodiments of any of the kits described herein can further include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) panels that can be secured over each of the one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) areas in the wall, such that leakage of the fluid from the interior of the pod to the exterior of the pod through the wall is prevented. The one or more panels can be made of one or more of any of the exemplary materials described herein that can be used to make the wall. The one or more panels are designed to cover any of the areas described herein. The one or more panels can be designed to click, snap, or screw into the wall to cover the one or more areas in the wall described herein.

Some embodiments of the kits described herein can further include an outer sleeve that can be secured over the pod in order to cover the at least one area in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented. As can be appreciated in the art, the sleeve can be composed, at least in part, of flexible rubber or silicon, such that it is capable of being slid over the container to cover the at least one area in the wall. In some embodiments, the sleeve can be an empty chromatography column that fits over the container.

Some embodiments of the kits described herein can further include a movable outer sleeve connected to the wall, such that rotation of the movable outer sleeve can cover the at least one area in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented. In some embodiments, the necessary rotation of the movable outer sleeve to cover the at least one area in the wall is clockwise. In some embodiments, the necessary rotation of the movable outer sleeve to cover the at least one area in the wall is counter-clockwise. In an alternative embodiment, a movable outer sleeve is connected to the wall, such that sliding the movable outer sleeve up or down can cover the at least one area in the wall, such that leakage of a fluid from the interior of the pod to the exterior of the pod through the at least one area in the wall is prevented.

Some embodiments of the kits further include an elution outlet that can be stably fixed onto the bottom end. The elution outlet can be a funnel or a flat surface having a port. In some embodiments, the elution outlet can include a means for controlling the flow of the fluid out of the interior of the container. Non-limiting examples of means for controlling the flow of a fluid out of the interior of the container include a screw valve or a stopcock. Additional exemplary means for controlling the flow of fluid out of the interior of the container are known in the art. In other examples, the elution outlet does not include a means for controlling the flow of a fluid out of the interior of the container. The elution outlet can be designed such that it can be snapped, clicked, or screwed onto the bottom end of the container. In some embodiments of any of the kits provided herein, the buffer reservoir can be sealed to the pod using one or more (e.g., two or three) O-rings (e.g., fixed to the pod or the buffer reservoir).

Some embodiments of any of the kits described herein further include a buffer reservoir that can be stably fixed onto the top end. The buffer reservoir can be snapped, clicked, or screwed onto the top end of the container. The buffer reservoir can have any shape such that it can hold a volume of fluid that can be flowed into the container. For example, a buffer reservoir can be cylindrical, rectangular, hemispherical, spherical, or ellipsoidal in shape. In some embodiments, the buffer reservoir can include a means for controlling the flow of the fluid into of the interior of the container from the buffer reservoir. Non-limiting examples of means for controlling the flow of a fluid out of the buffer reservoir into the container include a screw valve or a stopcock. Additional exemplary means for controlling the flow of fluid out of the buffer reservoir into the container are known in the art. In other examples, the buffer reservoir does not include a means for controlling the flow of a fluid out of the buffer reservoir into the interior of the container. The buffer reservoir can be designed to hold a volume of fluid between 0.1 mL to about 500 mL, about 0.1 mL to about 450 mL, about 0.1 mL to about 400 mL, about 0.1 mL to about 350 mL, about 0.1 mL to about 300 mL, about 0.1 mL to about 250 mL, about 0.1 mL to about 200 mL, about 0.1 mL to about 150 mL, about 0.1 mL to about 100 mL, about 0.1 mL to about 80 mL, about 0.1 mL to about 60 mL, about 0.1 mL to about 50 mL, about 0.1 mL to about 40 mL, about 0.1 mL to about 30 mL, about 0.1 mL to about 20 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 5 mL, about 0.1 mL to about 2 mL, about 0.1 mL to about 1 mL, about 0.2 mL to about 500 mL, about 0.2 mL to about 450 mL, about 0.2 mL to about 400 mL, about 0.2 mL to about 350 mL, about 0.2 mL to about 300 mL, about 0.2 mL to about 250 mL, about 0.2 mL to about 200 mL, about 0.2 mL to about 150 mL, about 0.2 mL to about 100 mL, about 0.2 mL to about 80 mL, about 0.2 mL to about 60 mL, about 0.2 mL to about 50 mL, about 0.2 mL to about 40 mL, about 0.2 mL to about 30 mL, about 0.2 mL to about 20 mL, about 0.2 mL to about 10 mL, about 0.2 mL to about 5 mL, about 0.2 mL to about 2 mL, about 0.2 mL to about 1 mL, 0.5 mL to about 500 mL, about 0.5 mL to about 450 mL, about 0.5 mL to about 400 mL, about 0.5 mL to about 350 mL, about 0.5 mL to about 300 mL, about 0.5 mL to about 250 mL, about 0.5 mL to about 200 mL, about 0.5 mL to about 150 mL, about 0.5 mL to about 100 mL, about 0.5 mL to about 80 mL, about 0.5 mL to about 60 mL, about 0.5 mL to about 50 mL, about 0.5 mL to about 40 mL, about 0.5 mL to about 30 mL, about 0.5 mL to about 20 mL, about 0.5 mL to about 10 mL, about 0.5 mL to about 5 mL, about 0.5 mL to about 2 mL, about 2 mL to about 500 mL, about 2 mL to about 450 mL, about 2 mL to about 400 mL, about 2 mL to about 350 mL, about 2 mL to about 300 mL, about 2 mL to about 250 mL, about 2 mL to about 200 mL, about 2 mL to about 150 mL, about 2 mL to about 100 mL, about 2 mL to about 80 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 40 mL, about 2 mL to about 30 mL, about 2 mL to about 20 mL, about 2 mL to about 10 mL, about 2 mL to about 5 mL, about 5 mL to about 500 mL, about 5 mL to about 450 mL, about 5 mL to about 400 mL, about 5 mL to about 350 mL, about 5 mL to about 300 mL, about 5 mL to about 250 mL, about 5 mL to about 200 mL, about 5 mL to about 150 mL, about 5 mL to about 100 mL, about 5 mL to about 80 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 40 mL, about 5 mL to about 30 mL, about 5 mL to about 20 mL, about 5 mL to about 10 mL, about 10 mL to about 500 mL, about 10 mL to about 450 mL, about 10 mL to about 400 mL, about 10 mL to about 350 mL, about 10 mL to about 300 mL, about 10 mL to about 250 mL, about 10 mL to about 200 mL, about 10 mL to about 150 mL, about 10 mL to about 100 mL, about 10 mL to about 80 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 40 mL, about 10 mL to about 30 mL, about 10 mL to about 20 mL, about 20 mL to about 500 mL, about 20 mL to about 450 mL, about 20 mL to about 400 mL, about 20 mL to about 350 mL, about 20 mL to about 300 mL, about 20 mL to about 250 mL, about 20 mL to about 200 mL, about 20 mL to about 150 mL, about 20 mL to about 100 mL, about 20 mL to about 80 mL, about 20 mL to about 60 mL, about 20 mL to about 50 mL, about 20 mL to about 40 mL, about 20 mL to about 30 mL, about 30 mL to about 500 mL, about 30 mL to about 450 mL, about 30 mL to about 400 mL, about 30 mL to about 350 mL, about 30 mL to about 300 mL, about 30 mL to about 250 mL, about 30 mL to about 200 mL, about 30 mL to about 150 mL, about 30 mL to about 100 mL, about 30 mL to about 80 mL, about 30 mL to about 60 mL, about 30 mL to about 50 mL, about 30 mL to about 40 mL, about 50 mL to about 500 mL, about 50 mL to about 450 mL, about 50 mL to about 400 mL, about 50 mL to about 350 mL, about 50 mL to about 300 mL, about 50 mL to about 250 mL, about 50 mL to about 200 mL, about 50 mL to about 150 mL, about 50 mL to about 100 mL, about 50 mL to about 80 mL, or about 50 mL to about 60 mL.

Also provided herein are kits that include: (a) any of the pods described herein (e.g., a pod having any of the characteristics or aspects described herein in any combination); (b) an inlet system having a top end, an elongated cylindrical body comprising a fluid conduit, and a bottom end, where the top end of the inlet system includes a fluid inlet and is configured for stable attachment to the top end of the pod and, when stably connected to the pod, allows for flow of a fluid into the pod through the fluid inlet and the fluid conduit, and out of the bottom end of the inlet system, and the elongated cylindrical body of the inlet system is configured to fit inside the wall of the pod; and (c) an outlet system including a fluid outlet that is configured for stable attachment to the bottom end of the pod, and when stably connected to the bottom end of the pod, allows for flow of a fluid out of the pod through the fluid outlet. In some embodiments of these kits, the top end of the inlet system and the top end of the pod include threading that allows for the stable attachment of the top end of the pod to the top end of the inlet system. In some embodiments of these kits, when the inlet system and the pod are stably attached, the top end of the inlet system and the top end of the pod form a seal e.g., a fluid-tight seal). In some embodiments of these kits, the threading in the top end of the inlet system or the threading in the top end of the pod includes an O-ring. Other means for providing a stable connection between the top end of the inlet system and the top end of the pod are known in the art. In some embodiments of these kits, the pod contains a chromatography resin; and the inlet system and the pod are configured such that when the inlet system and the pod are stably attached, the bottom end of the inlet system is proximal to the chromatography resin. In some embodiments of these kits, an interior surface of the pod and the bottom end of the inlet system are configured to allow for stable attachment of the pod and the bottom end of the inlet system. In some embodiments of these kits, when the inlet system and the pod are stably attached, the interior surface of the pod and the bottom end of the inlet system form a seal (e.g., a fluid-tight seal). In some embodiments of these kits, the interior surface of the pod and the bottom end of the inlet system include threading that allows for stable attachment of the pod and the bottom end of the inlet system. In some embodiments of these kits, the threading in the interior surface of the pod or the threading in the bottom end of the inlet system includes an O-ring. Other means for providing a stable connection between the interior surface of the pod and the bottom end of the inlet system are known in the art. In some embodiments of these kits, the outlet system and the bottom end of the pod include threading that allows for stable attachment of the outlet system to the bottom end of the pod. In some embodiments of these kits, the threading in the outlet system or the threading in the bottom end of the pod includes an O-ring. In some embodiments of these kits, the fluid inlet is capable of attachment to tubing (e.g., tubing which is connected to a chromatography system). In some embodiments of these kits, the fluid outlet is capable of attachment to tubing (e.g., tubing which is connected to a chromatography system). Some embodiments of these kits further include: (d) a means for stably positioning the long axis of the pod in a horizontal position in the interior of a culture vessel. Some embodiments of these kits further include: (d) a means for stably positioning the long axis of the pod in a vertical position in the interior of a culture vessel. In some embodiments of these kits, the elongated cylindrical body of the inlet system has a length of about 0.1 cm to about 50 cm, about 0.1 cm to about 45 cm, about 0.1 cm to about 40 cm, about 0.1 cm to about 35 cm, about 0.1 cm to about 30 cm, about 0.1 cm to about 25 cm, about 0.1 cm to about 20 cm, about 0.1 cm to about 18 cm, about 0.1 cm to about 16 cm, about 0.1 cm to about 14 cm, about 0.1 cm to about 12 cm, about 0.1 cm to about 10 cm, about 0.1 cm to about 8 cm, about 0.1 cm to about 6 cm, about 0.1 cm to about 5 cm, about 0.1 cm to about 4.5 cm, about 0.1 cm to about 4.0 cm, about 0.1 cm to about 3.5 cm, about 0.1 cm to about 3.0 cm, about 0.1 cm to about 2.5 cm, about 0.1 cm to about 2.0 cm, about 0.1 cm to about 1.5 cm, about 0.1 cm to about 1.0 cm, about 0.1 cm to about 0.5 cm, about 0.5 cm to about 50 cm, about 0.5 cm to about 45 cm, about 0.5 cm to about 40 cm, about 0.5 cm to about 35 cm, about 0.5 cm to about 30 cm, about 0.5 cm to about 25 cm, about 0.5 cm to about 20 cm, about 0.5 cm to about 18 cm, about 0.5 cm to about 16 cm, about 0.5 cm to about 14 cm, about 0.5 cm to about 12 cm, about 0.5 cm to about 10 cm, about 0.5 cm to about 8 cm, about 0.5 cm to about 6 cm, about 0.5 cm to about 5 cm, about 0.5 cm to about 4.5 cm, about 0.5 cm to about 4.0 cm, about 0.5 cm to about 3.5 cm, about 0.5 cm to about 3.0 cm, about 0.5 cm to about 2.5 cm, about 0.5 cm to about 2.0 cm, about 0.5 cm to about 1.5 cm, about 0.5 cm to about 1.0 cm, about 1.0 cm to about 50 cm, about 1.0 cm to about 45 cm, about 1.0 cm to about 40 cm, about 1.0 cm to about 35 cm, about 1.0 cm to about 30 cm, about 1.0 cm to about 25 cm, about 1.0 cm to about 20 cm, about 1.0 cm to about 18 cm, about 1.0 cm to about 16 cm, about 1.0 cm to about 14 cm, about 1.0 cm to about 12 cm, about 1.0 cm to about 10 cm, about 1.0 cm to about 8 cm, about 1.0 cm to about 6 cm, about 1.0 cm to about 5 cm, about 1.0 cm to about 4.5 cm, about 1.0 cm to about 4.0 cm, about 1.0 cm to about 3.5 cm, about 1.0 cm to about 3.0 cm, about 1.0 cm to about 2.5 cm, about 1.0 cm to about 2.0 cm, about 1.0 cm to about 1.5 cm, about 1.5 cm to about 50 cm, about 1.5 cm to about 45 cm, about 1.5 cm to about 40 cm, about 1.5 cm to about 35 cm, about 1.5 cm to about 30 cm, about 1.5 cm to about 25 cm, about 1.5 cm to about 20 cm, about 1.5 cm to about 18 cm, about 1.5 cm to about 16 cm, about 1.5 cm to about 14 cm, about 1.5 cm to about 12 cm, about 1.5 cm to about 10 cm, about 1.5 cm to about 8 cm, about 1.5 cm to about 6 cm, about 1.5 cm to about 5 cm, about 1.5 cm to about 4.5 cm, about 1.5 cm to about 4.0 cm, about 1.5 cm to about 3.5 cm, about 1.5 cm to about 3.0 cm, about 1.5 cm to about 2.5 cm, about 1.5 cm to about 2.0 cm, about 2.0 cm to about 50 cm, about 2.0 cm to about 45 cm, about 2.0 cm to about 40 cm, about 2.0 cm to about 35 cm, about 2.0 cm to about 30 cm, about 2.0 cm to about 25 cm, about 2.0 cm to about 20 cm, about 2.0 cm to about 18 cm, about 2.0 cm to about 16 cm, about 2.0 cm to about 14 cm, about 2.0 cm to about 12 cm, about 2.0 cm to about 10 cm, about 2.0 cm to about 8 cm, about 2.0 cm to about 6 cm, about 2.0 cm to about 5 cm, about 2.0 cm to about 4.5 cm, about 2.0 cm to about 4.0 cm, about 2.0 cm to about 3.5 cm, about 2.0 cm to about 3.0 cm, about 2.0 cm to about 2.5 cm, about 2.5 cm to about 50 cm, about 2.5 cm to about 45 cm, about 2.5 cm to about 40 cm, about 2.5 cm to about 35 cm, about 2.5 cm to about 30 cm, about 2.5 cm to about 25 cm, about 2.5 cm to about 20 cm, about 2.5 cm to about 18 cm, about 2.5 cm to about 16 cm, about 2.5 cm to about 14 cm, about 2.5 cm to about 12 cm, about 2.5 cm to about 10 cm, about 2.5 cm to about 8 cm, about 2.5 cm to about 6 cm, about 2.5 cm to about 5 cm, about 2.5 cm to about 4.5 cm, about 2.5 cm to about 4.0 cm, about 2.5 cm to about 3.5 cm, about 2.5 cm to about 3.0 cm, about 3.0 cm to about 50 cm, about 3.0 cm to about 45 cm, about 3.0 cm to about 40 cm, about 3.0 cm to about 35 cm, about 3.0 cm to about 30 cm, about 3.0 cm to about 25 cm, about 3.0 cm to about 20 cm, about 3.0 cm to about 18 cm, about 3.0 cm to about 16 cm, about 3.0 cm to about 14 cm, about 3.0 cm to about 12 cm, about 3.0 cm to about 10 cm, about 3.0 cm to about 8 cm, about 3.0 cm to about 6 cm, about 3.0 cm to about 5 cm, about 3.0 cm to about 4.5 cm, about 3.0 cm to about 4.0 cm, about 3.0 cm to about 3.5 cm, about 3.5 cm to about 50 cm, about 3.5 cm to about 45 cm, about 3.5 cm to about 40 cm, about 3.5 cm to about 35 cm, about 3.5 cm to about 30 cm, about 3.5 cm to about 25 cm, about 3.5 cm to about 20 cm, about 3.5 cm to about 18 cm, about 3.5 cm to about 16 cm, about 3.5 cm to about 14 cm, about 3.5 cm to about 12 cm, about 3.5 cm to about 10 cm, about 3.5 cm to about 8 cm, about 3.5 cm to about 6 cm, about 3.5 cm to about 5 cm, about 3.5 cm to about 4.5 cm, about 3.5 cm to about 4.0 cm, about 4.0 cm to about 50 cm, about 4.0 cm to about 45 cm, about 4.0 cm to about 40 cm, about 4.0 cm to about 35 cm, about 4.0 cm to about 30 cm, about 4.0 cm to about 25 cm, about 4.0 cm to about 20 cm, about 4.0 cm to about 18 cm, about 4.0 cm to about 16 cm, about 4.0 cm to about 14 cm, about 4.0 cm to about 12 cm, about 4.0 cm to about 10 cm, about 4.0 cm to about 8 cm, about 4.0 cm to about 6 cm, about 4.0 cm to about 5 cm, about 4.0 cm to about 4.5 cm, about 4.5 cm to about 50 cm, about 4.5 cm to about 45 cm, about 4.5 cm to about 40 cm, about 4.5 cm to about 35 cm, about 4.5 cm to about 30 cm, about 4.5 cm to about 25 cm, about 4.5 cm to about 20 cm, about 4.5 cm to about 18 cm, about 4.5 cm to about 16 cm, about 4.5 cm to about 14 cm, about 4.5 cm to about 12 cm, about 4.5 cm to about 10 cm, about 4.5 cm to about 8 cm, about 4.5 cm to about 6 cm, about 4.5 cm to about 5 cm, about 5 cm to about 50 cm, about 5 cm to about 45 cm, about 5 cm to about 40 cm, about 5 cm to about 35 cm, about 5 cm to about 30 cm, about 5 cm to about 25 cm, about 5 cm to about 20 cm, about 5 cm to about 18 cm, about 5 cm to about 16 cm, about 5 cm to about 14 cm, about 5 cm to about 12 cm, about 5 cm to about 10 cm, about 5 cm to about 8 cm, about 5 cm to about 6 cm, about 6 cm to about 50 cm, about 6 cm to about 45 cm, about 6 cm to about 40 cm, about 6 cm to about 35 cm, about 6 cm to about 30 cm, about 6 cm to about 25 cm, about 6 cm to about 20 cm, about 6 cm to about 18 cm, about 6 cm to about 16 cm, about 6 cm to about 14 cm, about 6 cm to about 12 cm, about 6 cm to about 10 cm, about 6 cm to about 8 cm, about 8 cm to about 50 cm, about 8 cm to about 45 cm, about 8 cm to about 40 cm, about 8 cm to about 35 cm, about 8 cm to about 30 cm, about 8 cm to about 25 cm, about 8 cm to about 20 cm, about 8 cm to about 18 cm, about 8 cm to about 16 cm, about 8 cm to about 14 cm, about 8 cm to about 12 cm, about 8 cm to about 10 cm, about 10 cm to about 50 cm, about 10 cm to about 45 cm, about 10 cm to about 40 cm, about 10 cm to about 35 cm, about 10 cm to about 30 cm, about 10 cm to about 25 cm, about 10 cm to about 20 cm, about 10 cm to about 18 cm, about 10 cm to about 16 cm, about 10 cm to about 14 cm, about 10 cm to about 12 cm, about 12 cm to about 50 cm, about 12 cm to about 45 cm, about 12 cm to about 40 cm, about 12 cm to about 35 cm, about 12 cm to about 30 cm, about 12 cm to about 25 cm, about 12 cm to about 20 cm, about 12 cm to about 18 cm, about 12 cm to about 16 cm, about 12 cm to about 14 cm, about 14 cm to about 50 cm, about 14 cm to about 45 cm, about 14 cm to about 40 cm, about 14 cm to about 35 cm, about 14 cm to about 30 cm, about 14 cm to about 25 cm, about 14 cm to about 20 cm, about 14 cm to about 18 cm, about 14 cm to about 16 cm, about 16 cm to about 50 cm, about 16 cm to about 45 cm, about 16 cm to about 40 cm, about 16 cm to about 35 cm, about 16 cm to about 30 cm, about 16 cm to about 25 cm, about 16 cm to about 20 cm, about 16 cm to about 18 cm, about 18 cm to about 50 cm, about 18 cm to about 45 cm, about 18 cm to about 40 cm, about 18 cm to about 35 cm, about 18 cm to about 30 cm, about 18 cm to about 25 cm, about 18 cm to about 20 cm, about 20 cm to about 50 cm, about 20 cm to about 45 cm, about 20 cm to about 40 cm, about 20 cm to about 35 cm, about 20 cm to about 30 cm, about 20 cm to about 25 cm, about 25 cm to about 50 cm, about 25 cm to about 45 cm, about 25 cm to about 40 cm, about 25 cm to about 35 cm, about 25 cm to about 30 cm, about 30 cm to about 50 cm, about 30 cm to about 45 cm, about 30 cm to about 40 cm, about 30 cm to about 35 cm, about 35 cm to about 50 cm, about 35 cm to about 45 cm, about 35 cm to about 40 cm, about 40 cm to about 50 cm, about 40 cm to about 45 cm, or about 45 cm to about 50 cm; and/or a diameter or an average diameter of about 0.1 cm to about 30 cm, about 0.1 cm to about 28 cm, about 0.1 cm to about 26 cm, about 0.1 cm to about 24 cm, about 0.1 cm to about 22 cm, about 0.1 cm to about 20 cm, about 0.1 cm to about 18 cm, about 0.1 cm to about 16 cm, about 0.1 cm to about 14 cm, about 0.1 cm to about 12 cm, about 0.1 cm to about 10 cm, about 0.1 cm to about 9 cm, about 0.1 cm to about 8 cm, about 0.1 cm to about 7 cm, about 0.1 cm to about 6 cm, about 0.1 cm to about 5 cm, about 0.1 cm to about 4 cm, about 0.1 cm to about 3 cm, about 0.1 cm to about 2 cm, about 0.1 cm to about 1 cm, about 0.1 cm to about 0.5 cm, about 0.5 cm to about 30 cm, about 0.5 cm to about 28 cm, about 0.5 cm to about 26 cm, about 0.5 cm to about 24 cm, about 0.5 cm to about 22 cm, about 0.5 cm to about 20 cm, about 0.5 cm to about 18 cm, about 0.5 cm to about 16 cm, about 0.5 cm to about 14 cm, about 0.5 cm to about 12 cm, about 0.5 cm to about 10 cm, about 0.5 cm to about 9 cm, about 0.5 cm to about 8 cm, about 0.5 cm to about 7 cm, about 0.5 cm to about 6 cm, about 0.5 cm to about 5 cm, about 0.5 cm to about 4 cm, about 0.5 cm to about 3 cm, about 0.5 cm to about 2 cm, about 0.5 cm to about 1 cm, about 1 cm to about 30 cm, about 1 cm to about 28 cm, about 1 cm to about 26 cm, about 1 cm to about 24 cm, about 1 cm to about 22 cm, about 1 cm to about 20 cm, about 1 cm to about 18 cm, about 1 cm to about 16 cm, about 1 cm to about 14 cm, about 1 cm to about 12 cm, about 1 cm to about 10 cm, about 1 cm to about 9 cm, about 1 cm to about 8 cm, about 1 cm to about 7 cm, about 1 cm to about 6 cm, about 1 cm to about 5 cm, about 1 cm to about 4 cm, about 1 cm to about 3 cm, about 1 cm to about 2 cm, about 2 cm to about 30 cm, about 2 cm to about 28 cm, about 2 cm to about 26 cm, about 2 cm to about 24 cm, about 2 cm to about 22 cm, about 2 cm to about 20 cm, about 2 cm to about 18 cm, about 2 cm to about 16 cm, about 2 cm to about 14 cm, about 2 cm to about 12 cm, about 2 cm to about 10 cm, about 2 cm to about 9 cm, about 2 cm to about 8 cm, about 2 cm to about 7 cm, about 2 cm to about 6 cm, about 2 cm to about 5 cm, about 2 cm to about 4 cm, about 2 cm to about 3 cm, about 3 cm to about 30 cm, about 3 cm to about 28 cm, about 3 cm to about 26 cm, about 3 cm to about 24 cm, about 3 cm to about 22 cm, about 3 cm to about 20 cm, about 3 cm to about 18 cm, about 3 cm to about 16 cm, about 3 cm to about 14 cm, about 3 cm to about 12 cm, about 3 cm to about 10 cm, about 3 cm to about 9 cm, about 3 cm to about 8 cm, about 3 cm to about 7 cm, about 3 cm to about 6 cm, about 3 cm to about 5 cm, about 3 cm to about 4 cm, about 4 cm to about 30 cm, about 4 cm to about 28 cm, about 4 cm to about 26 cm, about 4 cm to about 24 cm, about 4 cm to about 22 cm, about 4 cm to about 20 cm, about 4 cm to about 18 cm, about 4 cm to about 16 cm, about 4 cm to about 14 cm, about 4 cm to about 12 cm, about 4 cm to about 10 cm, about 4 cm to about 9 cm, about 4 cm to about 8 cm, about 4 cm to about 7 cm, about 4 cm to about 6 cm, about 4 cm to about 5 cm, about 5 cm to about 30 cm, about 5 cm to about 28 cm, about 5 cm to about 26 cm, about 5 cm to about 24 cm, about 5 cm to about 22 cm, about 5 cm to about 20 cm, about 5 cm to about 18 cm, about 5 cm to about 16 cm, about 5 cm to about 14 cm, about 5 cm to about 12 cm, about 5 cm to about 10 cm, about 5 cm to about 9 cm, about 5 cm to about 8 cm, about 5 cm to about 7 cm, about 5 cm to about 6 cm, about 6 cm to about 30 cm, about 6 cm to about 28 cm, about 6 cm to about 26 cm, about 6 cm to about 24 cm, about 6 cm to about 22 cm, about 6 cm to about 20 cm, about 6 cm to about 18 cm, about 6 cm to about 16 cm, about 6 cm to about 14 cm, about 6 cm to about 12 cm, about 6 cm to about 10 cm, about 6 cm to about 9 cm, about 6 cm to about 8 cm, about 6 cm to about 7 cm, about 7 cm to about 30 cm, about 7 cm to about 28 cm, about 7 cm to about 26 cm, about 7 cm to about 24 cm, about 7 cm to about 22 cm, about 7 cm to about 20 cm, about 7 cm to about 18 cm, about 7 cm to about 16 cm, about 7 cm to about 14 cm, about 7 cm to about 12 cm, about 7 cm to about 10 cm, about 7 cm to about 9 cm, about 7 cm to about 8 cm, about 8 cm to about 30 cm, about 8 cm to about 28 cm, about 8 cm to about 26 cm, about 8 cm to about 24 cm, about 8 cm to about 22 cm, about 8 cm to about 20 cm, about 8 cm to about 18 cm, about 8 cm to about 16 cm, about 8 cm to about 14 cm, about 8 cm to about 12 cm, about 8 cm to about 10 cm, about 8 cm to about 9 cm, about 9 cm to about 30 cm, about 9 cm to about 28 cm, about 9 cm to about 26 cm, about 9 cm to about 24 cm, about 9 cm to about 22 cm, about 9 cm to about 20 cm, about 9 cm to about 18 cm, about 9 cm to about 16 cm, about 9 cm to about 14 cm, about 9 cm to about 12 cm, about 9 cm to about 10 cm, about 10 cm to about 30 cm, about 10 cm to about 28 cm, about 10 cm to about 26 cm, about 10 cm to about 24 cm, about 10 cm to about 22 cm, about 10 cm to about 20 cm, about 10 cm to about 18 cm, about 10 cm to about 16 cm, about 10 cm to about 14 cm, about 10 cm to about 12 cm, about 12 cm to about 30 cm, about 12 cm to about 28 cm, about 12 cm to about 26 cm, about 12 cm to about 24 cm, about 12 cm to about 22 cm, about 12 cm to about 20 cm, about 12 cm to about 18 cm, about 12 cm to about 16 cm, about 12 cm to about 14 cm, about 14 cm to about 30 cm, about 14 cm to about 28 cm, about 14 cm to about 26 cm, about 14 cm to about 24 cm, about 14 cm to about 22 cm, about 14 cm to about 20 cm, about 14 cm to about 18 cm, about 14 cm to about 16 cm, about 16 cm to about 30 cm, about 16 cm to about 28 cm, about 16 cm to about 26 cm, about 16 cm to about 24 cm, about 16 cm to about 22 cm, about 16 cm to about 20 cm, about 16 cm to about 18 cm, about 18 cm to about 30 cm, about 18 cm to about 28 cm, about 18 cm to about 26 cm, about 18 cm to about 24 cm, about 18 cm to about 22 cm, about 18 cm to about 20 cm, about 20 cm to about 30 cm, about 20 cm to about 28 cm, about 20 cm to about 26 cm, about 20 cm to about 24 cm, about 20 cm to about 22 cm, about 22 cm to about 30 cm, about 22 cm to about 28 cm, about 22 cm to about 26 cm, about 22 cm to about 24 cm, about 24 cm to about 30 cm, about 24 cm to about 28 cm, about 24 cm to about 26 cm, about 26 cm to about 30 cm, about 26 cm to about 28 cm, or about 28 cm to about 30 cm.

In some embodiments, the fluid conduit in the inlet system has an internal diameter of about 0.1 mm to about 10 mm, about 0.1 mm to about 9.5 mm, about 0.1 mm to about 9.0 mm, about 0.1 mm to about 9.0 mm, about 0.1 mm to about 8.5 mm, about 0.1 mm to about 8.0 mm, about 0.1 mm to about 7.5 mm, about 0.1 mm to about 7.0 mm, about 0.1 mm to about 6.5 mm, about 0.1 mm to about 6.0 mm, about 0.1 mm to about 5.5 mm, about 0.1 mm to about 5.0 mm, about 0.1 mm to about 4.5 mm, about 0.1 mm to about 4.0 mm, about 0.1 mm to about 3.5 mm, about 0.1 mm to about 3.0 mm, about 0.1 mm to about 2.5 mm, about 0.1 mm to about 2.0 mm, about 0.1 mm to about 1.5 mm, about 0.1 mm to about 1.0 mm, about 0.1 mm to about 0.5 mm, about 0.5 mm to about 10 mm, about 0.5 mm to about 9.5 mm, about 0.5 mm to about 9.0 mm, about 0.5 mm to about 9.0 mm, about 0.5 mm to about 8.5 mm, about 0.5 mm to about 8.0 mm, about 0.5 mm to about 7.5 mm, about 0.5 mm to about 7.0 mm, about 0.5 mm to about 6.5 mm, about 0.5 mm to about 6.0 mm, about 0.5 mm to about 5.5 mm, about 0.5 mm to about 5.0 mm, about 0.5 mm to about 4.5 mm, about 0.5 mm to about 4.0 mm, about 0.5 mm to about 3.5 mm, about 0.5 mm to about 3.0 mm, about 0.5 mm to about 2.5 mm, about 0.5 mm to about 2.0 mm, about 0.5 mm to about 1.5 mm, about 0.5 mm to about 1.0 mm, about 1.0 mm to about 10 mm, about 1.0 mm to about 9.5 mm, about 1.0 mm to about 9.0 mm, about 1.0 mm to about 9.0 mm, about 1.0 mm to about 8.5 mm, about 1.0 mm to about 8.0 mm, about 1.0 mm to about 7.5 mm, about 1.0 mm to about 7.0 mm, about 1.0 mm to about 6.5 mm, about 1.0 mm to about 6.0 mm, about 1.0 mm to about 5.5 mm, about 1.0 mm to about 5.0 mm, about 1.0 mm to about 4.5 mm, about 1.0 mm to about 4.0 mm, about 1.0 mm to about 3.5 mm, about 1.0 mm to about 3.0 mm, about 1.0 mm to about 2.5 mm, about 1.0 mm to about 2.0 mm, or about 1.0 mm to about 1.5 mm.

In some embodiments of these kits, the pod contains a chromatography resin (e.g., any of the types of chromatography resin described herein), and the inlet system and the pod are configured such that when the inlet system and the pod are stably attached, the bottom end of the inlet system is approximately about 1 mm to about 30 mm (e.g., about 1 mm to about 25 mm, about 1 mm to about 20 mm, about 1 mm to about 15 mm, about 1 mm to about 10 mm, about 1 mm to about 5 mm) from the chromatography resin. In some embodiments of these kits, the pod can contain any of the volumes of chromatography resin described herein.

In some embodiments of these kits, the inlet system has an approximately conical shape. In some embodiments of these kits, the outlet system has an approximately conical shape. A non-limiting example of a kit including a pod, an inlet system, and an outlet system is shown in Figures 27 (non-assembled) and Figure 28 (assembled). Non-limiting examples of the other kits described herein are shown in Figures 1A-1E, Figure 3, Figures 23-26, and Figure 35.

Some embodiments of these kits can further include: (d) an adapter having a top end and a bottom end, wherein the top end of the adapter is configured for stable attachment to the bottom end of the pod, and the bottom end of the adapter has one or more blades, where when the top end of the adapter is stably attached to the bottom end of the pod, the positioning of the one or more blades promotes the flow of a fluid into the pod (e.g., through the material that allows for a liquid to reversibly flow from the exterior of the pod into the interior of the pod). A non-limiting example of such an adapter is shown in Figure 34. A diagram showing an exemplary adapter attached to an exemplary pod is shown in Figure 35.

In some embodiments of these kits, an interior surface of the pod and the top end of the adapter are configured to allow for stable attachment of the pod and the top end of the adapter. In some embodiments of these kits, when the adapter and the pod are stably attached, the interior surface of the pod and the top end of the adapter form a seal (e.g., a fluid-tight seal). In some embodiments of these kits, the interior surface of the pod and the top end of the adapter include threading that allows for stable attachment of the pod and the top end of the adapter. In some embodiments of these kits, the threading in the interior surface of the pod or the threading in the top end of the adapter includes an O-ring. Other means for providing a stable connection between the interior surface of the pod and the top end of the adapter are known in the art.

Some embodiments of any of the kits described herein can further include instructions for performing any of the methods described herein. Methods

Provided herein are methods of capturing a recombinant protein produced by a liquid cell culture that include: contacting any of the pods provided herein, with a liquid cell culture for a period of time, where the contacting step results in at least a portion of the at least one area contacting the liquid cell culture, and the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin.

Also provided herein are methods of purifying a recombinant protein produced by a liquid cell culture that include: (a) contacting any of the pods provided herein, with a liquid cell culture for a period of time, where the contacting step results in at least a portion of the at least one area contacting the liquid cell culture, and the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin; and (b) eluting the recombinant protein from the chromatography resin after the period of time, thereby purifying the recombinant protein. In some embodiments, the eluting (and optional washing step) can be performed using a chromatography system (e.g., an ÄKTA system).

In some embodiments of any of the methods described herein, the pod is any of the pods described herein, where the pod includes a chromatography resin (e.g., any of the chromatography resins described herein or known in the art) disposed in the interior of the container. In some embodiments, the pod is disposed such that the longitudinal axis of the pod is placed about horizontally (e.g., horizontally) in a liquid cell culture. In some embodiments, the pod is disposed such that the longitudinal axis of the pod is placed about vertically (e.g., vertically) in a liquid cell culture. In some embodiments, the pod includes a chromatography resin that is a commercial chromatography resin. In some embodiments, the chromatography resin is an affinity chromatography resin (e.g., a protein A resin, a protein G resin, a protein L resin, a nickel resin, or a magnetic bead resin). In some embodiments, after the contacting step, the pod or the chromatography resin can be frozen or stored for a period of time before the recombinant protein is eluted from the chromatography resin in the pod. In some embodiments where the pod or chromatography resin is frozen or stored for a period of time, the pod can or chromatography resin can be frozen or stored for a period of time after the contacting step and after the pod or chromatography resin has been contacted with a wash buffer. In some embodiments, the contacting results in at least a portion of the at least one area (e.g., at least 10% of the at least one area, at least 15% of the at least one area, at least 20% of the at least one area, at least 25% of the at least one area, at least 30% of the at least one area, at least 35% of the at least one area, at least 40% of the at least one area, at least 45% of the at least one area, at least 50% of the at least one area, at least 55% of the at least one area, at least 60% of the at least one area, at least 65% of the at least one area, at least 70% of the at least one area, at least 75% of the at least one area, at least 80% of the at least one area, at least 85% of the at least one area, at least 90% of the at least one area, at least 95% of the at least one area, at least 97% of the at least one area, at least 98% of the at least one area, at least 99% of the at least one area) contacting the liquid cell culture. In some embodiments, at least 20% (e.g., at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) of the total surface area of the at least one area is in contact with the liquid culture.

In some embodiments, the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin. In some embodiments, the capturing can result in the recombinant protein being bound to a level of up to 60% to 100% (e.g., up to 60% to 70%, up to 60% to 75%, up to 60% to 80%, up to 60% to 85%, up to 60% to 90%, up to 60% to 95%, up to 65% to 70%, up to 65% to 75%, up to 65% to 80%, up to 65% to 85%, up to 65% to 90%, up to 65% to 95%, up to 65% to 100%, up to 70% to 75%, up to 70% to 80%, up to 70% to 85%, up to 70% to 90%, up to 70% to 95%, up to 70% to 100%, up to 75% to 80%, up to 75% to 85%, up to 75% to 90%, up to 75% to 95%, up to 75% to 100%, up to 80% to 85%, up to 80% to 90%, up to 80% to 95%, up to 80% to 100%, up to 85% to 90%, up to 85% to 95%, up to 85% to 100%, up to 90% to 95%, up to 95% to 100%, up to 50% to 55%, up to 50% to 60%, up to 50% to 65%, up to 50% to 70%, up to 50% to 75%, up to 50% to 80%, up to 50% to 85%, up to 50% to 90%, up to 50% to 95%, up to 50% to 100%, up to 55% to 60%, up to 55% to 65%, up to 55% to 70%, up to 55% to 75%, up to 55% to 80%, up to 55% to 85%, up to 55% to 90%, up to 55% to 95%, up to 55% to 100%, up to 50%, up to 51%, up to 52%, up to 53%, up to 54%, up to 55%, up to 56%, up to 57%, up to 58%, up to 59%, up to 60%, up to 61%, up to 62%, up to 63%, up to 64%, up to 65%, up to 66%, up to 67%, up to 68%, up to 69%, up to 70%, up to 71%, up to 72%, up to 73%, up to 74%, up to 75%, up to 76%, up to 77%, up to 78%, up to 79%, up to 80%, up to 81%, up to 82%, up to 83%, up to 84%, up to 85%, up to 86%, up to 87%, up to 88%, up to 89%, up to 90%, up to 91%, up to 92%, up to 93%, up to 94%, up to 95%, up to 96%, up to 97%, up to 98%, up to 99%, or 100%) of the chromatography resin binding capacity.

In some embodiments, the capturing can result in the capture of about 0.05 mg to about 80 mg (e.g., about 0.05 mg to about 0.1 mg, about 0.05 mg to about 0.2 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.6 mg, about 0.05 mg to about 0.8 mg, about 0.05 mg to about 1 mg, about 0.05 mg to about 2 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 15 mg, about 0.05 mg to about 20 mg, about 0.05 mg to about 25 mg, about 0.05 mg to about 30 mg, about 0.05 mg to about 35 mg, about 0.05 mg to about 40 mg, about 0.05 mg to about 45 mg, about 0.05 mg to about 50 mg, about 0.05 mg to about 55 mg, about 0.05 mg to about 60 mg, about 0.05 mg to about 65 mg, about 0.05 mg to about 70 mg, about 0.05 mg to about 75 mg, about 0.05 mg to about 80 mg, about 0.1 mg to about 0.2 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.6 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 1 mg, about 0.1 mg to about 2 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 10 mg, about 0.1 mg to about 15 mg, about 0.1 mg to about 20 mg, about 0.1 mg to about 25 mg, about 0.1 mg to about 30 mg, about 0.1 mg to about 35 mg, about 0.1 mg to about 40 mg, about 0.1 mg to about 45 mg, about 0.1 mg to about 50 mg, about 0.1 mg to about 55 mg, about 0.1 mg to about 60 mg, about 0.1 mg to about 65 mg, about 0.1 mg to about 70 mg, about 0.1 mg to about 75 mg, about 0.1 mg to about 80 mg, about 0.2 mg to about 0.5 mg, about 0.2 mg to about 0.6 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 1 mg, about 0.2 mg to about 2 mg, about 0.2 mg to about 5 mg, about 0.2 mg to about 10 mg, about 0.2 mg to about 15 mg, about 0.2 mg to about 20 mg, about 0.2 mg to about 25 mg, about 0.2 mg to about 30 mg, about 0.2 mg to about 35 mg, about 0.2 mg to about 40 mg, about 0.2 mg to about 45 mg, about 0.2 mg to about 50 mg, about 0.2 mg to about 55 mg, about 0.2 mg to about 60 mg, about 0.2 mg to about 65 mg, about 0.2 mg to about 70 mg, about 0.2 mg to about 75 mg, about 0.2 mg to about 80 mg, about 0.5 mg to about 0.6 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 1 mg, about 0.5 mg to about 2 mg, about 0.5 mg to about 5 mg, about 0.5 mg to about 10 mg, about 0.5 mg to about 15 mg, about 0.5 mg to about 20 mg, about 0.5 mg to about 25 mg, about 0.5 mg to about 30 mg, about 0.5 mg to about 35 mg, about 0.5 mg to about 40 mg, about 0.5 mg to about 45 mg, about 0.5 mg to about 50 mg, about 0.5 mg to about 55 mg, about 0.5 mg to about 60 mg, about 0.5 mg to about 65 mg, about 0.5 mg to about 70 mg, about 0.5 mg to about 75 mg, about 0.5 mg to about 80 mg, about 0.6 mg to about 0.8 mg, about 0.6 mg to about 1 mg, about 0.6 mg to about 2 mg, about 0.6 mg to about 5 mg, about 0.6 mg to about 10 mg, about 0.6 mg to about 15 mg, about 0.6 mg to about 20 mg, about 0.6 mg to about 25 mg, about 0.6 mg to about 30 mg, about 0.6 mg to about 35 mg, about 0.6 mg to about 40 mg, about 0.6 mg to about 45 mg, about 0.6 mg to about 50 mg, about 0.6 mg to about 55 mg, about 0.6 mg to about 60 mg, about 0.6 mg to about 65 mg, about 0.6 mg to about 70 mg, about 0.6 mg to about 75 mg, about 0.6 mg to about 80 mg, about 0.8 mg to about 1 mg, about 0.8 mg to about 2 mg, about 0.8 mg to about 5 mg, about 0.8 mg to about 10 mg, about 0.8 mg to about 15 mg, about 0.8 mg to about 20 mg, about 0.8 mg to about 25 mg, about 0.8 mg to about 30 mg, about 0.8 mg to about 35 mg, about 0.8 mg to about 40 mg, about 0.8 mg to about 45 mg, about 0.8 mg to about 50 mg, about 0.8 mg to about 55 mg, about 0.8 mg to about 60 mg, about 0.8 mg to about 65 mg, about 0.8 mg to about 70 mg, about 0.8 mg to about 75 mg, about 0.8 mg to about 80 mg, about 1 mg to about 2 mg, about 1 mg to about 5 mg, about 1 mg to about 10 mg, about 1 mg to about 15 mg, about 1 mg to about 20 mg, about 1 mg to about 25 mg, about 1 mg to about 30 mg, about 1 mg to about 35 mg, about 1 mg to about 40 mg, about 1 mg to about 45 mg, about 1 mg to about 50 mg, about 1 mg to about 55 mg, about 1 mg to about 60 mg, about 1 mg to about 65 mg, about 1 mg to about 70 mg, about 1 mg to about 75 mg, about 1 mg to about 80 mg, about 2 mg to about 5 mg, about 2 mg to about 10 mg, about 2 mg to about 15 mg, about 2 mg to about 20 mg, about 2 mg to about 25 mg, about 2 mg to about 30 mg, about 2 mg to about 35 mg, about 2 mg to about 40 mg, about 2 mg to about 45 mg, about 2 mg to about 50 mg, about 2 mg to about 55 mg, about 2 mg to about 60 mg, about 2 mg to about 65 mg, about 2 mg to about 70 mg, about 2 mg to about 75 mg, about 2 mg to about 80 mg, about 5 mg to about 10 mg, about 5 mg to about 15 mg, about 5 mg to about 20 mg, about 5 mg to about 25 mg, about 5 mg to about 30 mg, about 5 mg to about 35 mg, about 5 mg to about 40 mg, about 5 mg to about 45 mg, about 5 mg to about 50 mg, about 5 mg to about 55 mg, about 5 mg to about 60 mg, about 5 mg to about 65 mg, about 5 mg to about 70 mg, about 5 mg to about 75 mg, about 5 mg to about 80 mg, about 10 mg to about 15 mg, about 10 mg to about 20 mg, about 10 mg to about 25 mg, about 10 mg to about 30 mg, about 10 mg to about 35 mg, about 10 mg to about 40 mg, about 10 mg to about 45 mg, about 10 mg to about 50 mg, about 10 mg to about 55 mg, about 10 mg to about 60 mg, about 10 mg to about 65 mg, about 10 mg to about 70 mg, about 10 mg to about 75 mg, about 10 mg to about 80 mg, about 15 mg to about 20 mg, about 15 mg to about 20 mg, about 15 mg to about 25 mg, about 15 mg to about 30 mg, about 15 mg to about 35 mg, about 15 mg to about 40 mg, about 15 mg to about 45 mg, about 15 mg to about 50 mg, about 15 mg to about 55 mg, about 15 mg to about 60 mg, about 15 mg to about 65 mg, about 15 mg to about 70 mg, about 15 mg to about 75 mg, about 15 mg to about 80 mg, about 20 mg to about 25 mg, about 20 mg to about 30 mg, about 20 mg to about 35 mg, about 20 mg to about 40 mg, about 20 mg to about 45 mg, about 20 mg to about 50 mg, about 20 mg to about 55 mg, about 20 mg to about 60 mg, about 20 mg to about 65 mg, about 20 mg to about 70 mg, about 20 mg to about 75 mg, about 20 mg to about 80 mg, about 25 mg to about 30 mg, about 25 mg to about 35 mg, about 25 mg to about 40 mg, about 25 mg to about 45 mg, about 25 mg to about 50 mg, about 25 mg to about 55 mg, about 25 mg to about 60 mg, about 25 mg to about 65 mg, about 25 mg to about 70 mg, about 25 mg to about 75 mg, about 25 mg to about 80 mg, about 30 mg to about 35 mg, about 30 mg to about 40 mg, about 30 mg to about 45 mg, about 30 mg to about 50 mg, about 30 mg to about 55 mg, about 30 mg to about 60 mg, about 30 mg to about 65 mg, about 30 mg to about 70 mg, about 30 mg to about 75 mg, about 30 mg to about 80 mg, about 35 mg to about 40 mg, about 35 mg to about 45 mg, about 35 mg to about 50 mg, about 35 mg to about 55 mg, about 35 mg to about 60 mg, about 35 mg to about 65 mg, about 35 mg to about 70 mg, about 35 mg to about 75 mg, about 35 mg to about 80 mg, about 40 mg to about 45 mg, about 40 mg to about 50 mg, about 45 mg to about 55 mg, about 45 mg to about 60 mg, about 45 mg to about 65 mg, about 45 mg to about 70 mg, about 45 mg to about 75 mg, about 45 mg to about 80 mg, about 50 mg to about 55 mg, about 50 mg to about 60 mg, about 50 mg to about 65 mg, about 50 mg to about 70 mg, about 50 mg to about 75 mg, about 50 mg to about 80 mg, about 55 mg to about 60 mg, about 55 mg to about 65 mg, about 55 mg to about 70 mg, about 55 mg to about 75 mg, about 55 mg to about 80 mg, about 60 mg to about 65 mg, about 60 mg to about 70 mg, about 60 mg to about 75 mg, about 60 mg to about 80 mg, about 65 mg to about 70 mg, about 65 mg to about 75 mg, about 65 mg to about 80 mg, about 70 mg to about 75 mg, about 70 mg to about 80 mg, about 75 mg to about 80 mg) of recombinant protein on the chromatography resin.

In some embodiments, the purity of capture is at least 45% (e.g., at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.2%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%) as determined by analytical size exclusion chromatography (SEC). Period of Time

In some embodiments of any of the methods provided herein, the contacting of any of the pods provided herein with a liquid cell culture is for a period of time (e.g., about 30 seconds to about 2 weeks). In some embodiments of any of the methods provided herein, the eluting of the recombinant protein from the chromatography resin is after a period of time (e.g., about 30 seconds to about 2 weeks).

In some embodiments, the period of time for contacting and/or for eluting the recombinant protein from the chromatography resin is about 30 seconds to about 1 minute, about 30 seconds to 45 seconds, about 30 seconds to about 90 seconds, about 30 seconds to about 2 minutes, about 30 seconds to about 150 seconds, about 30 seconds to about 3 minutes, about 30 seconds to about 210 seconds, about 30 seconds to about 4 minutes, about 30 seconds to about 270 seconds, about 30 seconds to about 5 minutes, about 1 minute to about 2 minutes, about 1 minute to about 3 minutes, about 1 minute to about 4 minutes, about 1 minute to about 5 minutes, about 2 minutes to about 3 minutes, about 2 minutes to about 4 minutes, about 2 to about 5 minutes, about 3 minutes to about 4 minutes, about 3 minutes to about 5 minutes, about 4 minutes to about 5 minutes, about 5 minutes to about 2 weeks, about 5 minutes to about 168 hours, about 5 minutes to about 162 hours, about 5 minutes to about 156 hours, about 5 minutes to about 150 hours, about 5 minutes to about 144 hours, about 5 minutes to about 138 hours, about 5 minutes to about 132 hours, about 5 minutes to about 126 hours, about 5 minutes to about 120 hours, about 5 minutes to about 114 hours, about 5 minutes to about 108 hours, about 5 minutes to about 102 hours, about 5 minutes to about 96 hours, about 5 minutes to about 90 hours, about 5 minutes to about 84 hours, about 5 minutes to about 78 hours, about 5 minutes to about 72 hours, about 5 minutes to about 66 hours, about 5 minutes to about 60 hours, about 5 minutes to about 54 hours, about 5 minutes to about 48hours, about 5 minutes to about 42 hours, about 5 minutes to about 36 hours, about 5 minutes to about 30 hours, about 5 minutes to about 24 hours, about 5 minutes to about 18 hours, about 5 minutes to about 12 hours, about 5 minutes to about 6 hours, about 5 minutes to about 5 hours, about 5 minutes to about 240 minutes, about 5 minutes to about 210 minutes, about 5 minutes to about 180 minutes, about 5 minutes to about 150 minutes, about 5 minutes to about 120 minutes, about 5 minutes to about 90 minutes, about 5 minutes to about 60 minutes, about 5 minutes to about 30 minutes, about 5 minutes to about 20 minutes, about 5 minutes to about 15 minutes, about 5 minutes to about 10 minutes; about 10 minutes to about 2 weeks, about 10 minutes to about 168 hours, about 10 minutes to about 162 hours, about 10 minutes to about 156 hours, about 10 minutes to about 150 hours, about 10 minutes to about 144 hours, about 10 minutes to about 138 hours, about 10 minutes to about 132 hours, about 10 minutes to about 126 hours, about 10 minutes to about 120 hours, about 10 minutes to about 114 hours, about 10 minutes to about 108 hours, about 10 minutes to about 102 hours, about 10 minutes to about 96 hours, about 10 minutes to about 90 hours, about 10 minutes to about 84 hours, about 10 minutes to about 78 hours, about 10 minutes to about 72 hours, about 10 minutes to about 66 hours, about 10 minutes to about 60 hours, about 10 minutes to about 54 hours, about 10 minutes to about 48hours, about 10 minutes to about 42 hours, about 10 minutes to about 36 hours, about 10 minutes to about 30 hours, about 10 minutes to about 24 hours, about 10 minutes to about 18 hours, about 10 minutes to about 12 hours, about 10 minutes to about 6 hours, about 10 minutes to about 5 hours, about 10 minutes to about 240 minutes, about 10 minutes to about 210 minutes, about 10 minutes to about 180 minutes, about 10 minutes to about 150 minutes, about 10 minutes to about 120 minutes, about 10 minutes to about 90 minutes, about 10 minutes to about 60 minutes, about 10 minutes to about 30 minutes, about 10 minutes to about 15 minutes, about 10 minutes to about 20 minutes, about 10 minutes to about 25 minutes; about 20 minutes to about 2 weeks, about 20 minutes to about 168 hours, about 20 minutes to about 162 hours, about 20 minutes to about 156 hours, about 20 minutes to about 150 hours, about 20 minutes to about 144 hours, about 20 minutes to about 138 hours, about 20 minutes to about 132 hours, about 20 minutes to about 126 hours, about 20 minutes to about 120 hours, about 20 minutes to about 114 hours, about 20 minutes to about 108 hours, about 20 minutes to about 102 hours, about 20 minutes to about 96 hours, about 20 minutes to about 90 hours, about 20 minutes to about 84 hours, about 20 minutes to about 78 hours, about 20 minutes to about 72 hours, about 20 minutes to about 66 hours, about 20 minutes to about 60 hours, about 20 minutes to about 54 hours, about 20 minutes to about 48hours, about 20 minutes to about 42 hours, about 20 minutes to about 36 hours, about 20 minutes to about 30 hours, about 20 minutes to about 24 hours, about 20 minutes to about 18 hours, about 20 minutes to about 12 hours, about 20 minutes to about 6 hours, about 20 minutes to about 5 hours, about 20 minutes to about 240 minutes, about 20 minutes to about 210 minutes, about 20 minutes to about 180 minutes, about 20 minutes to about 150 minutes, about 20 minutes to about 120 minutes, about 20 minutes to about 90 minutes, about 20 minutes to about 60 minutes, about 20 minutes to about 30 minutes, about 20 minutes to about 25 minutes; about 30 minutes to about 2 weeks, about 30 minutes to about 168 hours, about 30 minutes to about 162 hours, about 30 minutes to about 156 hours, about 30 minutes to about 150 hours, about 30 minutes to about 144 hours, about 30 minutes to about 138 hours, about 30 minutes to about 132 hours, about 30 minutes to about 126 hours, about 30 minutes to about 120 hours, about 30 minutes to about 114 hours, about 30 minutes to about 108 hours, about 30 minutes to about 102 hours, about 30 minutes to about 96 hours, about 30 minutes to about 90 hours, about 30 minutes to about 84 hours, about 30 minutes to about 78 hours, about 30 minutes to about 72 hours, about 30 minutes to about 66 hours, about 30 minutes to about 60 hours, about 30 minutes to about 54 hours, about 30 minutes to about 48hours, about 30 minutes to about 42 hours, about 30 minutes to about 36 hours, about 30 minutes to about 30 hours, about 30 minutes to about 24 hours, about 30 minutes to about 18 hours, about 30 minutes to about 12 hours, about 30 minutes to about 6 hours, about 30 minutes to about 5 hours, about 30 minutes to about 240 minutes, about 30 minutes to about 210 minutes, about 30 minutes to about 180 minutes, about 30 minutes to about 150 minutes, about 30 minutes to about 120 minutes, about 30 minutes to about 90 minutes, about 30 minutes to about 60 minutes; about 40 minutes to about 2 weeks, about 40 minutes to about 168 hours, about 40 minutes to about 162 hours, about 40 minutes to about 156 hours, about 40 minutes to about 150 hours, about 40 minutes to about 144 hours, about 40 minutes to about 138 hours, about 40 minutes to about 132 hours, about 40 minutes to about 126 hours, about 40 minutes to about 120 hours, about 40 minutes to about 114 hours, about 40 minutes to about 108 hours, about 40 minutes to about 102 hours, about 40 minutes to about 96 hours, about 40 minutes to about 90 hours, about 40 minutes to about 84 hours, about 40 minutes to about 78 hours, about 40 minutes to about 72 hours, about 40 minutes to about 66 hours, about 40 minutes to about 60 hours, about 40 minutes to about 54 hours, about 40 minutes to about 48hours, about 40 minutes to about 42 hours, about 40 minutes to about 36 hours, about 40 minutes to about 30 hours, about 40 minutes to about 24 hours, about 40 minutes to about 18 hours, about 40 minutes to about 12 hours, about 40 minutes to about 6 hours, about 40 minutes to about 5 hours, about 40 minutes to about 240 minutes, about 40 minutes to about 210 minutes, about 40 minutes to about 180 minutes, about 40 minutes to about 150 minutes, about 40 minutes to about 120 minutes, about 40 minutes to about 90 minutes, about 40 minutes to about 60 minutes, about 40 minutes to about 50 minutes, about 40 minutes to about 45 minutes; about 45 minutes to about 2 weeks, about 45 minutes to about 168 hours, about 45 minutes to about 162 hours, about 45 minutes to about 156 hours, about 45 minutes to about 150 hours, about 45 minutes to about 144 hours, about 45 minutes to about 138 hours, about 45 minutes to about 132 hours, about 45 minutes to about 126 hours, about 45 minutes to about 120 hours, about 45 minutes to about 114 hours, about 45 minutes to about 108 hours, about 45 minutes to about 102 hours, about 45 minutes to about 96 hours, about 45 minutes to about 90 hours, about 45 minutes to about 84 hours, about 45 minutes to about 78 hours, about 45 minutes to about 72 hours, about 45 minutes to about 66 hours, about 45 minutes to about 60 hours, about 45 minutes to about 54 hours, about 45 minutes to about 48hours, about 45 minutes to about 42 hours, about 45 minutes to about 36 hours, about 45 minutes to about 30 hours, about 45 minutes to about 24 hours, about 45 minutes to about 18 hours, about 45 minutes to about 12 hours, about 45 minutes to about 6 hours, about 45 minutes to about 5 hours, about 45 minutes to about 240 minutes, about 45 minutes to about 210 minutes, about 45 minutes to about 180 minutes, about 45 minutes to about 150 minutes, about 45 minutes to about 120 minutes, about 45 minutes to about 90 minutes, about 45 minutes to about 60 minutes, about 45 minutes to about 50 minutes, about 45 minutes to about 55 minutes; about 60 minutes to about 2 weeks, about 60 minutes to about 168 hours, about 60 minutes to about 162 hours, about 60 minutes to about 156 hours, about 60 minutes to about 150 hours, about 60 minutes to about 144 hours, about 60 minutes to about 138 hours, about 60 minutes to about 132 hours, about 60 minutes to about 126 hours, about 60 minutes to about 120 hours, about 60 minutes to about 114 hours, about 60 minutes to about 108 hours, about 60 minutes to about 102 hours, about 60 minutes to about 96 hours, about 60 minutes to about 90 hours, about 60 minutes to about 84 hours, about 60 minutes to about 78 hours, about 60 minutes to about 72 hours, about 60 minutes to about 66 hours, about 60 minutes to about 60 hours, about 60 minutes to about 54 hours, about 60 minutes to about 48hours, about 60 minutes to about 42 hours, about 60 minutes to about 36 hours, about 60 minutes to about 30 hours, about 60 minutes to about 24 hours, about 60 minutes to about 18 hours, about 60 minutes to about 12 hours, about 60 minutes to about 6 hours, about 60 minutes to about 5 hours, about 60 minutes to about 240 minutes, about 60 minutes to about 210 minutes, about 60 minutes to about 180 minutes, about 60 minutes to about 150 minutes, about 60 minutes to about 120 minutes, about 60 minutes to about 90 minutes, about 60 minutes to about 85 minutes, about 60 minutes to about 80 minutes, about 60 minutes to about 75 minutes, about 60 minutes to about 70 minutes; about 70 minutes to about 2 weeks, about 70 minutes to about 168 hours, about 70 minutes to about 162 hours, about 70 minutes to about 156 hours, about 70 minutes to about 150 hours, about 70 minutes to about 144 hours, about 70 minutes to about 138 hours, about 70 minutes to about 132 hours, about 70 minutes to about 126 hours, about 70 minutes to about 120 hours, about 70 minutes to about 114 hours, about 70 minutes to about 108 hours, about 70 minutes to about 102 hours, about 70 minutes to about 96 hours, about 70 minutes to about 90 hours, about 70 minutes to about 84 hours, about 70 minutes to about 78 hours, about 70 minutes to about 72 hours, about 70 minutes to about 66 hours, about 70 minutes to about 60 hours, about 70 minutes to about 54 hours, about 70 minutes to about 48hours, about 70 minutes to about 42 hours, about 70 minutes to about 36 hours, about 70 minutes to about 30 hours, about 70 minutes to about 24 hours, about 70 minutes to about 18 hours, about 70 minutes to about 12 hours, about 70 minutes to about 6 hours, about 70 minutes to about 5 hours, about 70 minutes to about 240 minutes, about 70 minutes to about 210 minutes, about 70 minutes to about 180 minutes, about 70 minutes to about 150 minutes, about 70 minutes to about 120 minutes, about 70 minutes to about 90 minutes, about 70 minutes to about 85 minutes, about 70 minutes to about 80 minutes; about 75 minutes to about 80 minutes, about 75 minutes to about 85 minutes, about 75 minutes to about 90 minutes; about 90 minutes to about 2 weeks, about 90 minutes to about 168 hours, about 90 minutes to about 162 hours, about 90 minutes to about 156 hours, about 90 minutes to about 150 hours, about 90 minutes to about 144 hours, about 90 minutes to about 138 hours, about 90 minutes to about 132 hours, about 90 minutes to about 126 hours, about 90 minutes to about 120 hours, about 90 minutes to about 114 hours, about 90 minutes to about 108 hours, about 90 minutes to about 102 hours, about 90 minutes to about 96 hours, about 90 minutes to about 90 hours, about 90 minutes to about 84 hours, about 90 minutes to about 78 hours, about 90 minutes to about 72 hours, about 90 minutes to about 66 hours, about 90 minutes to about 60 hours, about 90 minutes to about 54 hours, about 90 minutes to about 48hours, about 90 minutes to about 42 hours, about 90 minutes to about 36 hours, about 90 minutes to about 30 hours, about 90 minutes to about 24 hours, about 90 minutes to about 18 hours, about 90 minutes to about 12 hours, about 90 minutes to about 6 hours, about 90 minutes to about 5 hours, about 90 minutes to about 240 minutes, about 90 minutes to about 210 minutes, about 90 minutes to about 180 minutes, about 90 minutes to about 150 minutes, about 90 minutes to about 120 minutes, about 90 minutes to about 100 minutes, about 90 minutes to about 110 minutes, about 100 minutes to about 110 minutes, about 100 minutes to about 120 minutes; about 120 minutes to about 2 weeks, about 120 minutes to about 168 hours, about 120 minutes to about 162 hours, about 120 minutes to about 156 hours, about 120 minutes to about 150 hours, about 120 minutes to about 144 hours, about 120 minutes to about 138 hours, about 120 minutes to about 132 hours, about 120 minutes to about 126 hours, about 120 minutes to about 120 hours, about 120 minutes to about 114 hours, about 120 minutes to about 108 hours, about 120 minutes to about 102 hours, about 120 minutes to about 96 hours, about 120 minutes to about 90 hours, about 120 minutes to about 84 hours, about 120 minutes to about 78 hours, about 120 minutes to about 72 hours, about 120 minutes to about 66 hours, about 120 minutes to about 60 hours, about 120 minutes to about 54 hours, about 120 minutes to about 48hours, about 120 minutes to about 42 hours, about 120 minutes to about 36 hours, about 120 minutes to about 30 hours, about 120 minutes to about 24 hours, about 120 minutes to about 18 hours, about 120 minutes to about 12 hours, about 120 minutes to about 6 hours, about 120 minutes to about 5 hours, about 120 minutes to about 240 minutes, about 120 minutes to about 210 minutes, about 120 minutes to about 180 minutes, about 120 minutes to about 150 minutes, about 120 minutes to about 130 minutes, about 120 minutes to about 140 minutes, about 130 minutes to about 140 minutes, about 130 minutes to about 150 minutes, about 140 minutes to about 150 minutes; about 150 minutes to about 2 weeks, about 150 minutes to about 168 hours, about 150 minutes to about 162 hours, about 150 minutes to about 156 hours, about 150 minutes to about 150 hours, about 150 minutes to about 144 hours, about 150 minutes to about 138 hours, about 150 minutes to about 132 hours, about 150 minutes to about 126 hours, about 150 minutes to about 120 hours, about 150 minutes to about 114 hours, about 150 minutes to about 108 hours, about 150 minutes to about 102 hours, about 150 minutes to about 96 hours, about 150 minutes to about 90 hours, about 150 minutes to about 84 hours, about 150 minutes to about 78 hours, about 150 minutes to about 72 hours, about 150 minutes to about 66 hours, about 150 minutes to about 60 hours, about 150 minutes to about 54 hours, about 150 minutes to about 48hours, about 150 minutes to about 42 hours, about 150 minutes to about 36 hours, about 150 minutes to about 30 hours, about 150 minutes to about 24 hours, about 150 minutes to about 18 hours, about 150 minutes to about 12 hours, about 150 minutes to about 6 hours, about 150 minutes to about 5 hours, about 150 minutes to about 240 minutes, about 150 minutes to about 210 minutes, about 150 minutes to about 180 minutes; about 180 minutes to about 2 weeks, about 180 minutes to about 168 hours, about 180 minutes to about 162 hours, about 180 minutes to about 156 hours, about 180 minutes to about 150 hours, about 180 minutes to about 144 hours, about 180 minutes to about 138 hours, about 180 minutes to about 132 hours, about 180 minutes to about 126 hours, about 180 minutes to about 120 hours, about 180 minutes to about 114 hours, about 180 minutes to about 108 hours, about 180 minutes to about 102 hours, about 180 minutes to about 96 hours, about 180 minutes to about 90 hours, about 180 minutes to about 84 hours, about 180 minutes to about 78 hours, about 180 minutes to about 72 hours, about 180 minutes to about 66 hours, about 180 minutes to about 60 hours, about 180 minutes to about 54 hours, about 180 minutes to about 48hours, about 180 minutes to about 42 hours, about 180 minutes to about 36 hours, about 180 minutes to about 30 hours, about 180 minutes to about 24 hours, about 180 minutes to about 18 hours, about 180 minutes to about 12 hours, about 180 minutes to about 6 hours, about 180 minutes to about 5 hours, about 180 minutes to about 240 minutes, about 180 minutes to about 210 minutes, about 180 minutes to about 200 minutes, about 180 minutes to about 190 minutes, about 190 minutes to about 200 minutes, about 190 minutes to about 210 minutes; about 210 minutes to about 2 weeks, about 210 minutes to about 168 hours, about 210 minutes to about 162 hours, about 210 minutes to about 156 hours, about 210 minutes to about 150 hours, about 210 minutes to about 144 hours, about 210 minutes to about 138 hours, about 210 minutes to about 132 hours, about 210 minutes to about 126 hours, about 210 minutes to about 120 hours, about 210 minutes to about 114 hours, about 210 minutes to about 108 hours, about 210 minutes to about 102 hours, about 210 minutes to about 96 hours, about 210 minutes to about 90 hours, about 210 minutes to about 84 hours, about 210 minutes to about 78 hours, about 210 minutes to about 72 hours, about 210 minutes to about 66 hours, about 210 minutes to about 60 hours, about 210 minutes to about 54 hours, about 210 minutes to about 48hours, about 210 minutes to about 42 hours, about 210 minutes to about 36 hours, about 210 minutes to about 30 hours, about 210 minutes to about 24 hours, about 210 minutes to about 18 hours, about 210 minutes to about 12 hours, about 210 minutes to about 6 hours, about 210 minutes to about 5 hours, about 210 minutes to about 240 minutes, about 210 minutes to about 230 minutes, about 210 minutes to about 220 minutes; about 240 minutes to about 2 weeks, about 240 minutes to about 168 hours, about 240 minutes to about 162 hours, about 240 minutes to about 156 hours, about 240 minutes to about 150 hours, about 240 minutes to about 144 hours, about 240 minutes to about 138 hours, about 240 minutes to about 132 hours, about 240 minutes to about 126 hours, about 240 minutes to about 120 hours, about 240 minutes to about 114 hours, about 240 minutes to about 108 hours, about 240 minutes to about 102 hours, about 240 minutes to about 96 hours, about 240 minutes to about 90 hours, about 240 minutes to about 84 hours, about 240 minutes to about 78 hours, about 240 minutes to about 72 hours, about 240 minutes to about 66 hours, about 240 minutes to about 60 hours, about 240 minutes to about 54 hours, about 240 minutes to about 48hours, about 240 minutes to about 42 hours, about 240 minutes to about 36 hours, about 240 minutes to about 30 hours, about 240 minutes to about 24 hours, about 240 minutes to about 18 hours, about 240 minutes to about 12 hours, about 240 minutes to about 6 hours, about 240 minutes to about 5 hours, about 240 minutes to about 280 minutes, about 240 minutes to about 270 minutes, about 240 minutes to about 260 minutes, about 240 minutes to about 250 minutes; about 270 minutes to about 280 minutes, about 270 minutes to about 290 minutes, about 270 minutes to about 5 hours; about 5 hours to about 2 weeks, about 5 hours to about 168 hours, about 5 hours to about 162 hours, about 5 hours to about 156 hours, about 5 hours to about 150 hours, about 5 hours to about 144 hours, about 5 hours to about 138 hours, about 5 hours to about 132 hours, about 5 hours to about 126 hours, about 5 hours to about 120 hours, about 5 hours to about 114 hours, about 5 hours to about 108 hours, about 5 hours to about 102 hours, about 5 hours to about 96 hours, about 5 hours to about 90 hours, about 5 hours to about 84 hours, about 5 hours to about 78 hours, about 5 hours to about 72 hours, about 5 hours to about 66 hours, about 5 hours to about 60 hours, about 5 hours to about 54 hours, about 5 hours to about 48hours, about 5 hours to about 42 hours, about 5 hours to about 36 hours, about 5 hours to about 30 hours, about 5 hours to about 24 hours, about 5 hours to about 18 hours, about 5 hours to about 12 hours, about 5 hours to about 11 hours, about 5 hours to about 10 hours, about 5 hours to about 9 hours, about 5 hours to about 8 hours, about 5 hours to about 7 hours, about 5 hours to about 6 hours; about 6 hours to about 12 hours, about 6 hours to about 8 hours, about 6 hours to about 10 hours, about 8 hours to about 10 hours, about 8 hours to about 12 hours; about 8 hours to about 2 weeks, about 8 hours to about 168 hours, about 8 hours to about 162 hours, about 6 hours to about 156 hours, about 6 hours to about 150 hours, about 6 hours to about 144 hours, about 6 hours to about 138 hours, about 6 hours to about 132 hours, about 6 hours to about 126 hours, about 6 hours to about 120 hours, about 6 hours to about 114 hours, about 6 hours to about 108 hours, about 6 hours to about 102 hours, about 6 hours to about 96 hours, about 6 hours to about 90 hours, about 6 hours to about 84 hours, about 6 hours to about 78 hours, about 6 hours to about 72 hours, about 6 hours to about 66 hours, about 6 hours to about 60 hours, about 6 hours to about 54 hours, about 6 hours to about 48hours, about 6 hours to about 42 hours, about 6 hours to about 36 hours, about 6 hours to about 30 hours, about 6 hours to about 24 hours, about 6 hours to about 18 hours, about 6 hours to about 12 hours, about 6 hours to about 11 hours, about 6 hours to about 10 hours, about 6 hours to about 9 hours, about 6 hours to about 8 hours, about 6 hours to about 7 hours, about 7 hours to about 8 hours; about 12 hours to about 2 weeks, about 12 hours to about 168 hours, about 12 hours to about 162 hours, about 12 hours to about 156 hours, about 12 hours to about 150 hours, about 12 hours to about 144 hours, about 12 hours to about 138 hours, about 12 hours to about 132 hours, about 12 hours to about 126 hours, about 12 hours to about 120 hours, about 12 hours to about 114 hours, about 12 hours to about 108 hours, about 12 hours to about 102 hours, about 12 hours to about 96 hours, about 12 hours to about 90 hours, about 12 hours to about 84 hours, about 12 hours to about 78 hours, about 12 hours to about 72 hours, about 12 hours to about 66 hours, about 12 hours to about 60 hours, about 12 hours to about 54 hours, about 12 hours to about 48hours, about 12 hours to about 42 hours, about 12 hours to about 36 hours, about 12 hours to about 30 hours, about 12 hours to about 24 hours, about 12 hours to about 18 hours, about 12 hours to about 14 hours, about 12 hours to about 16 hours, about 14 hours to about 16 hours, about 14 hours to about 18 hours, about 14 hours to about 24 hours, about 14 hours to about 36 hours, about 14 hours to about 48 hours, about 14 hours to about 36 hours, about 16 hours to about 18 hours, about 16 hours to about 20 hours, about 16 hours to about 24 hours, about 16 hours to about 36 hours, about 16 hours to about 48 hours, about 18 hours to about 24 hours, about 18 hours to about 36 hours, about 18 hours to about 48 hours; about 24 hours to about 2 weeks, about 24 hours to about 168 hours, about 24 hours to about 162 hours, about 24 hours to about 156 hours, about 24 hours to about 150 hours, about 24 hours to about 144 hours, about 24 hours to about 138 hours, about 24 hours to about 132 hours, about 24 hours to about 126 hours, about 24 hours to about 120 hours, about 24 hours to about 114 hours, about 24 hours to about 108 hours, about 24 hours to about 102 hours, about 24 hours to about 96 hours, about 24 hours to about 90 hours, about 24 hours to about 84 hours, about 24 hours to about 78 hours, about 24 hours to about 72 hours, about 24 hours to about 66 hours, about 24 hours to about 60 hours, about 24 hours to about 54 hours, about 24 hours to about 48hours, about 24 hours to about 42 hours, about 24 hours to about 36 hours, about 24 hours to about 30 hours, about 30 hours to about 36 hours; about 26 hours to about 28 hours, about 26 hours to about 30 hours, about 26 hours to about 36 hours, about 26 hours to about 48 hours, about 28 hours to about 30 hours, about 28 hours to about 36 hours, about 28 hours to about 48 hours, about 30 hours to about 36 hours, about 30 hours to about 48 hours; about 36 hours to about 2 weeks, about 36 hours to about 168 hours, about 36 hours to about 162 hours, about 36 hours to about 156 hours, about 36 hours to about 150 hours, about 36 hours to about 144 hours, about 36 hours to about 138 hours, about 36 hours to about 132 hours, about 36 hours to about 126 hours, about 36 hours to about 120 hours, about 36 hours to about 114 hours, about 36 hours to about 108 hours, about 36 hours to about 102 hours, about 36 hours to about 96 hours, about 36 hours to about 90 hours, about 36 hours to about 84 hours, about 36 hours to about 78 hours, about 36 hours to about 72 hours, about 36 hours to about 66 hours, about 36 hours to about 60 hours, about 36 hours to about 54 hours, about 36 hours to about 48 hours, about 36 hours to about 42 hours, about 36 hours to about 40 hours, about 40 hours to about 42 hours; about 48 hours to about 2 weeks, about 48 hours to about 168 hours, about 48 hours to about 162 hours, about 48 hours to about 156 hours, about 48 hours to about 150 hours, about 48 hours to about 144 hours, about 48 hours to about 138 hours, about 48 hours to about 132 hours, about 48 hours to about 126 hours, about 48 hours to about 120 hours, about 48 hours to about 114 hours, about 48 hours to about 108 hours, about 48 hours to about 102 hours, about 48 hours to about 96 hours, about 48 hours to about 90 hours, about 48 hours to about 84 hours, about 48 hours to about 78 hours, about 48 hours to about 72 hours, about 48 hours to about 66 hours, about 48 hours to about 60 hours, about 48 hours to about 54 hours; about 60 hours to about 2 weeks, about 60 hours to about 168 hours, about 60 hours to about 162 hours, about 60 hours to about 156 hours, about 60 hours to about 150 hours, about 60 hours to about 144 hours, about 60 hours to about 138 hours, about 60 hours to about 132 hours, about 60 hours to about 126 hours, about 60 hours to about 120 hours, about 60 hours to about 114 hours, about 60 hours to about 108 hours, about 60 hours to about 102 hours, about 60 hours to about 96 hours, about 60 hours to about 90 hours, about 60 hours to about 84 hours, about 60 hours to about 78 hours, about 60 hours to about 72 hours, about 60 hours to about 66 hours; about 72 hours to about 2 weeks, about 72 hours to about 168 hours, about 72 hours to about 162 hours, about 72 hours to about 156 hours, about 72 hours to about 150 hours, about 72 hours to about 144 hours, about 72 hours to about 138 hours, about 72 hours to about 132 hours, about 72 hours to about 126 hours, about 72 hours to about 120 hours, about 72 hours to about 114 hours, about 72 hours to about 108 hours, about 72 hours to about 102 hours, about 72 hours to about 96 hours, about 72 hours to about 90 hours, about 72 hours to about 84 hours, about 72 hours to about 78 hours; about 84 hours to about 2 weeks, about 84 hours to about 168 hours, about 84 hours to about 162 hours, about 84 hours to about 156 hours, about 84 hours to about 150 hours, about 84 hours to about 144 hours, about 84 hours to about 138 hours, about 84 hours to about 132 hours, about 84 hours to about 126 hours, about 84 hours to about 120 hours, about 84 hours to about 114 hours, about 84 hours to about 108 hours, about 84 hours to about 102 hours, about 84 hours to about 96 hours, about 84 hours to about 90 hours; about 96 hours to about 2 weeks, about 96 hours to about 168 hours, about 96 hours to about 162 hours, about 96 hours to about 156 hours, about 96 hours to about 150 hours, about 96 hours to about 144 hours, about 96 hours to about 138 hours, about 96 hours to about 132 hours, about 96 hours to about 126 hours, about 96 hours to about 120 hours, about 96 hours to about 114 hours, about 96 hours to about 108 hours, about 96 hours to about 102 hours; about 108 hours to about 2 weeks, about 108 hours to about 168 hours, about 108 hours to about 162 hours, about 108 hours to about 156 hours, about 108 hours to about 150 hours, about 108 hours to about 144 hours, about 108 hours to about 138 hours, about 108 hours to about 132 hours, about 108 hours to about 126 hours, about 108 hours to about 120 hours, about 108 hours to about 114 hours; about 120 hours to about 2 weeks, about 120 hours to about 168 hours, about 120 hours to about 162 hours, about 120 hours to about 156 hours, about 120 hours to about 150 hours, about 120 hours to about 144 hours, about 120 hours to about 138 hours, about 120 hours to about 132 hours, about 120 hours to about 126 hours; about 132 hours to about 2 weeks, about 132 hours to about 168 hours, about 132 hours to about 162 hours, about 132 hours to about 156 hours, about 132 hours to about 150 hours, about 132 hours to about 144 hours, about 132 hours to about 138 hours; about 144 hours to about 2 weeks, about 144 hours to about 168 hours, about 144 hours to about 162 hours, about 144 hours to about 156 hours, about 144 hours to about 150 hours; about 156 hours to about 2 weeks, about 156 hours to about 168 hours, about 156 hours to about 162 hours; or about 168 hours to about 2 weeks.

In some embodiments of the contacting step of any of the methods provided herein, the period of time occurs when the liquid cell culture is in its exponential growth phase. In some embodiments of the contacting step of any of the methods provided herein, the period of time occurs when the liquid cell culture is in its stationary growth phase. In some embodiments, the contacting can occur when a liquid cell culture is in its death phase. In some embodiments of the contacting step of any of the methods provided herein, the period of time occurs on day 4 of cell culture until day 5.

In some embodiments of any of the methods described herein, the contacting step can include contacting one or more pods (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) sequentially or at the same time with a liquid cell culture for a period of time (e.g., any of the period of times described herein). Recombinant Proteins

Recombinant proteins described herein include, without limitation, receptors, antibodies, antigen-binding antibody fragments, antibody fragments, fusion proteins, ligands, secreted proteins, enzymes, recombinant protein drug products, and any recombinant protein with or without a tag. Non-limiting examples of tags include: a fluorescent protein (e.g., green fluorescent protein, red fluorescent protein, cyan fluorescent protein, yellow fluorescent protein), a radioisotope, a His-tag, a FLAG- tag, a c-myc tag, a maltose-binding protein (MBP) tag, an albumin-binding protein (ABP) tag, a bacteriophage T7 epitope (T7-tag), a biotin-carboxy carrier protein (BCCP) tag, a bluetongue virus tag (B-tag), a calmodulin binding peptide tag, a cellulose binding domain tag, a chitin binding domain tag, a choline binding domain tag, a galactose-binding protein tag, a Glu-Glu epitope tag, a human influenza hemagglutinin tag, a Halo Tag®, a histidine affinity tag (HAT), a polyarginine tag (Asp-tag), a polyphenylalanine tag (Phe-tag), an S-TAG, a streptavidin-binding peptide, a streptavidin tag, a Strep-tag, an enzyme (e.g., alkaline phosphatase, chloramphenicol acetyl transferase (CAT), glutathione S-transferase). In some embodiments, the recombinant protein is a multimeric protein (e.g., a heterodimeric protein, a homodimeric protein, or a trimeric protein).

Exemplary recombinant antibodies and recombinant antibody fragments that can be expressed in any of the cells described herein can include recombinant IgG1 (e.g., recombinant human IgG1), recombinant IgG2 (e.g., recombinant human IgG2), recombinant IgG3 (e.g., recombinant human IgG3), recombinant IgG4 (e.g., recombinant human IgG4), recombinant IgM (e.g., recombinant human IgM), recombinant IgE (e.g., recombinant human IgE), recombinant IgD (e.g., recombinant human IgD), recombinant IgA (e.g., recombinant human IgA), a recombinant Fab, a recombinant scFv, a recombinant VNAR, a recombinant VHH, a recombinant diabody (e.g., a recombinant bispecific diabody), a recombinant scFv-Fc, a recombinant IgNAR, a recombinant bispecific diabody, a recombinant light chain variable domain, a recombinant heavy chain variable domain, a recombinant triabody, a recombinant tetrabody, a recombinant tandem scFv, a recombinant tandem scFv-Fc, a recombine, ant scFv-Fc knobs-into-holes, a recombinant scFv-Fc-scFv, a recombinant F(abʹ) ₂ , a recombinant Fab-scFv, a recombinant (FabʹscFv) ₂ , a recombinant scDiabody-Fc, a recombinant scDiabody, a recombinant scDiabody- CH3, a recombinant IgG-scFv, a recombinant V-IgG, a recombinant IgG-V, a recombinant two-in-one IgG, a recombinant dual V domain IgG, and a recombinant knobs-into-holes with a common light chain. In some embodiments, the recombinant protein is an antibody. As used herein, the term“antibody” refers to a whole or intact antibody (e.g., IgM, IgG, IgA, IgD, or IgE) molecule. An antibody is tetrameric in nature, containing two identical heavy chain protein portions and two identical light chain protein portions, which associate together to adopt a“Y”-shaped conformation held together by disulfide bridges at the hinge region. The term“antibody” includes a monoclonal antibody, a chimerized or chimeric antibody, a humanized antibody, a deimmunized human antibody, and a fully human antibody. The antibody can be made in or derived from any of a variety of species, e.g., mammals such as humans, non-human primates (e.g., monkeys, baboons, or chimpanzees), horses, cattle, pigs, sheep, goats, dogs, cats, rabbits, guinea pigs, gerbils, hamsters, rats, and mice. In some embodiments, the antibodies can be generated synthetically, e.g., via synthetically produced oligonucleotides, as described in, e.g, Fuh et al., J. Mol. Biol.340:1073-1093, 2004, and Knappik et al., J. Mol. Biol.296(11):57-86, 2000, the disclosures of each of which are incorporated herein by reference in their entirety. In some embodiments, the antibody can be generated by antibody phage display in yeast and/or human cells.

As used herein, the term“antibody fragment,”“antigen-binding fragment,” or similar terms refer to a fragment of an antibody that retains the ability to bind to an antigen, e.g., a single chain antibody (scFv), an Fd fragment, an Fab fragment, an Fab′ fragment, or an F(ab′)2 fragment. An scFv is a single polypeptide chain that includes both the heavy and light chain variable regions of the antibody from which the scFv is derived. In addition, diabodies (Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999, the disclosures of both of which are incorporated herein by reference in their entirety), minibodies, triabodies

(Schoonooghe et al., BMC Biotechnol.9:70, 2009), domain antibodies (also known as “heavy chain immunoglobulins” or camelids; Holt et al., Trends Biotechnol.

21(11):484-490, 2003); and intrabodies (Huston et al., Human Antibodies 10(3- 4):127-142, 2001; Wheeler et al., Mol. Ther.8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22): 960-966, 2004, the disclosures of each of which are incorporated herein by reference in their entirety) are included in the definition of antibody fragments and can be incorporated into the compositions, and used in the methods, described herein. In some embodiments, the recombinant protein is a bispecific antibody. Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens.

Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chain/light-chain pairs have different specificities (Milstein and Cuello, Nature 305:537-539, 1983). Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion of the heavy chain variable region is preferably with an immunoglobulin heavy-chain constant domain, including at least part of the hinge, CH2, and CH3 regions. For further details of illustrative currently known methods for generating bispecific antibodies see, e.g., Suresh et al., Methods Enzymol.121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985;

Shalaby et al., J. Exp. Med.175:217-225, 1992; Kostelny et al., J. Immunol.

148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A.90:6444-6448, 1993; Gruber et al., J. Immunol.152:5368, 1994; and Tutt et al., J. Immunol.147:60, 1991.

Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. See, e.g., Kostelny et al., J. Immunol.148(5):1547-1553, 1992. The leucine zipper peptides from the Fos and Jun proteins may be linked to the Fab′ portions of two different antibodies by gene fusion. The antibody homodimers may be reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The“diabody” technology described by Hollinger et al., Proc. Natl. Acad. Sci. U.S.A.90:6444-6448, 1993, has provided an alternative mechanism for making bispecific antibody fragments. The fragments include a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (scFv) dimers has also been reported. See, e.g., Gruber et al., J. Immunol.152:5368, 1994. Alternatively, the antibodies can be“linear antibodies” as described in, e.g., Zapata et al., Protein Eng.8(10):1057-1062, 1995. Briefly, these antibodies include a pair of tandem Fd segments (VH-CH1-VH-CH1) which form a pair of antigen binding regions. Linear antibodies can be bispecific or monospecific.

Antibodies with more than two valencies (e.g., trispecific antibodies) are contemplated and described in, e.g., Tutt et al., J. Immunol.147:60, 1991.

The disclosure also embraces variant forms of multi-specific antibodies such as the dual variable domain immunoglobulin (DVD-Ig) molecules described in Wu et al., Nat Biotechnol.25(11): 1290-1297, 2007. The DVD-Ig molecules are designed such that two different light chain variable domains (VL) from two different parent antibodies are linked in tandem directly or via a short linker by recombinant DNA techniques, followed by the light chain constant domain. Similarly, the heavy chain includes two different heavy chain variable domains (VH) linked in tandem, followed by the constant domain CH1 and Fc region. Methods for making DVD-Ig molecules from two parent antibodies are further described in, e.g., WO 08/024188 and WO 07/024715. In some embodiments, the bispecific antibody is a Fabs-in-Tandem immunoglobulin, in which the light chain variable region with a second specificity is fused to the heavy chain variable region of a whole antibody. Such antibodies are described in, e.g., WO 2015/103072.

In some embodiments, the recombinant protein is a nanobody, such as a camelid or dromedary antibodies (e.g., antibodies derived from Camelus bactrianus, Calelus dromaderius, or Lama paccos). Such antibodies, unlike the typical two-chain (fragment) or four-chain (whole antibody) antibodies from most mammals, generally lack light chains. See U.S. Patent No.5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem.14:440-448, 2003. As with other antibodies of non-human origin, an amino acid sequence of a camelid antibody can be altered recombinantly to obtain a sequence that more closely resembles a human sequence, i.e., the nanobody can be“humanized” to thereby further reduce the potential immunogenicity of the antibody.

In some embodiments, the recombinant protein is a non-antibody, scaffold protein. These proteins are, generally, obtained through combinatorial chemistry- based adaptation of preexisting antigen-binding proteins. For example, the binding site of human transferrin for human transferrin receptor can be diversified using the system described herein to create a diverse library of transferrin variants, some of which have acquired affinity for different antigens. Ali et al., J. Biol. Chem.

274:24066-24073, 1999. The portion of human transferrin not involved with binding the receptor remains unchanged and serves as a scaffold, like framework regions of antibodies, to present the variant binding sites.

One of skill in the art would appreciate that the scaffold portion of the non- antibody scaffold protein can include, e.g., all or part of: the Z domain of S. aureus protein A, human transferrin, human tenth fibronectin type III domain, kunitz domain of a human trypsin inhibitor, human CTLA-4, an ankyrin repeat protein, a human lipocalin (e.g., anticalins), human crystallin, human ubiquitin, or a trypsin inhibitor from E. elaterium.

In some embodiments, the recombinant protein is a fusion protein. Non- limiting examples of fusion proteins that can be produced by any of the cells described herein include chimeric antibodies (e.g., humanized antibodies), labelled antibodies (e.g., antibodies labelled with an enzyme or fluorescent protein tag), proteins ligands that bind specifically to a cancer cell surface receptor, where the protein ligand is labelled with a proteinaceous cytotoxin, an antibody with a fusion partner that extends the half life of the antibody in vivo (e.g., an antibody fused to human albumin), a bispecific antibody, where one light chain variable domain and one heavy chain variable domain bind to an antigen on an immune cell (e.g., a T cell, a macrophage, a neutrophil, or a monocyte) and the other light chain variable domain and the other heavy chain variable domain bind to an antigen on a target cell (e.g., a cancer cell or a virus infected cell) or a pathogen (e.g., a virus, a mycobacterium, or a parasite).

Further examples of recombinant proteins include, without limitation, ligands, enzymes, peptides, scFvs, non-antibody scaffold proteins, chimeric antigen receptor molecules, and the like, in which the diversity element is fused in frame with one or more constant elements. In some embodiments, the recombinant protein is a multimeric protein, such as, but not limited to, Fabs, whole antibodies, MHC class I molecules, MHC class I molecules, T cell receptor molecules, bispecific antibodies, trispecific antibodies, integrins, tumor growth factor receptor dimers, G-protein coupled receptors, and the like. Cells

The liquid cell culture can include any cell, e.g., a eukaryotic cell. As used herein, the term“eukaryotic cell” refers to a cell having a distinct, membrane-bound nucleus. Such cells may include, for example, mammalian (e.g., rodent, non-human primate, or human), insect, fungal, or plant cells. In some embodiments, the eukaryotic cell is a yeast cell, such as Saccharomyces cerevisiae. In some embodiments, the eukaryotic cell is a higher eukaryote, such as mammalian, avian, plant, or insect cells.

Non-limiting examples of mammalian cells that can produce a recombinant protein include, e.g., rodent (e.g., mouse, rat, or hamster) or human cells. In some embodiments, the mammalian cell is a Chinese hamster ovarian (CHO) cell, a human retina PER.C6 cell, a human aminocyte CAP cell, a mouse melanoma NS0 or Sp2/0 cell, a duck EB66 cell, a hamster BHK cell, a human embryonic kidney (HEK 293) cell, a COS cell, or a chicken-derived B cell line DT40. In some embodiments, the HEK 293 cell is a freestyle HEK293-F cell, a HEK 2936F, a HEK293FT, a HEK 293T cell, or an Expi293F cell. CHO cells are epithelial cells which grow as an adherent monolayer or in suspension. CHO cells lines that were derived from the original cells isolated by Tjio and Puck (Tjio et al., J. Exp. Med.108: 259-271, 1958) may include point mutations, deletions, or integrations of nucleic acid sequences relative to the original CHO cell lines. The subgroups of CHO cells may include, but is not limited to, CHO DP-12 cells, CHO-K1 cells, CHO/dhfr- cells, CHO-S cells, CHO-GS cells, CHO-K1 DUX B11 cells, CHO pro3- cells, CHO- DG44 cells, CHO GAT- cells, or freestyle CHO-S cells. In some embodiments, the eukaryotic cell is a primary cell or a cell line. In some embodiments, the mammalian cell is a cancer cell. In some embodiments, the mammalian cell is a human cancer cell line, including those listed in the“Cancer Cell Line Encyclopedia” (Barretina et al., Nature 483(7391):603-607, 2012, or Forbes et al., Curr. Protoc. Hum. Genet.2008). In some embodiments, the eukaryotic cell is a T lymphocyte lineage cell (e.g., a primary T cell or a T-cell line), a T-cell derived cell line which lacks TCR expression (see, e.g., Torres et al., J. Immunol.2003), or B lymphocyte lineage cell, such as pro-B cells (see, e.g., Urashima et al., Am. J. Hematol.1994), pre-B cells (see, e.g., Adachi et al., DNA Cell Biol.2006), and B-cells, and cell lines derived from these. In some embodiments, the B lineage cell line is a murine pre-B cell line, such as 1624-5 (Holms et al., J. Exp. Med.1986; Principato et al., Mol. Cell Biol.1990), or a pro-B cell line, such as Ba/F3 (commercially available from Thermo Scientific). The mammalian cells can express a secreted version of the recombinant protein. In some embodiments, the secreted version of the recombinant protein is transiently expressed. In some embodiments, the secreted version of the recombinant protein is stably expressed.

In some embodiments, the liquid cell culture can include an insect cell (e.g., a baculovirus-insect cell, a Drosophila cell (e.g., a Drosophila S2 cell), a Trichoplusia ni cell, a Lepidopteran cell), a yeast cell (e.g., a Saccharoymyces cerevisiae cell, a Pichia pastoris cell), a plant cell or a bacterial cell (e.g., an E.coli cell, a Baccillus brevis cell, a Bacillus megaterium cell, a Bacillus subtilis cell, a Caulobacter crescentis cell).

Skilled practitioners will appreciate that the proteins and vectors described herein can be introduced into the cells (e.g., mammalian cells, insect cells, yeast cells, bacterial cells, or plant cells). Non-limiting examples of vectors and methods for introducing vectors into cells are described herein. Methods of Culturing Cells

Methods of culturing cells are well known in the art. Cells can be maintained in vitro under conditions that favor growth, proliferation, continued viability, and differentiation. Generally speaking, cells can be cultured by contacting a cell with a liquid culture medium that includes the necessary growth factors and supplements to support cell viability and growth. The term“liquid culture medium” means a fluid that contains sufficient nutrients to allow a cell (e.g., any of the cells described herein) to grow or proliferate in vitro. For example, a liquid culture medium can contain one or more of: amino acids (e.g., 20 amino acids), a purine (e.g., hypoxanthine), a pyrimidine (e.g., thymidine), choline, inositol, thiamine, folic acid, biotin, calcium, niacinamide, pyridoxine, riboflavin, thymidine, cyanocobalamin, pyruvate, lipoic acid, magnesium, glucose, sodium, potassium, iron, copper, zinc, and sodium bicarbonate.

In some embodiments, a liquid culture medium can contain serum from a mammal. In some embodiments, a liquid culture medium does not contain serum or another extract from a mammal (a defined liquid culture medium). In some embodiments, a liquid culture medium can contain trace metals, a mammalian growth hormone, and/or a mammalian growth factor. Another example of liquid culture medium is minimal medium (e.g., a medium containing only inorganic salts, a carbon source, and water). Non-limiting examples of liquid culture medium are described herein. Additional examples of liquid culture medium are known in the art and are commercially available. A liquid cell culture medium can contain any density of any cell described herein (e.g., any density of mammalian cells).

As used herein, the term“animal-derived component free liquid culture medium” means a liquid culture medium that does not contain any components (e.g., proteins or serum) derived from a mammal. As used herein, the term“serum-free liquid culture medium” means a liquid culture medium that does not contain a mammalian serum. As used herein, the term“serum-containing liquid culture medium” means a liquid culture medium that contains a mammalian serum. As used herein, the term“chemically-defined liquid culture medium” is a term of art and means a liquid culture medium in which all of the chemical components are known. For example, a chemically-defined liquid culture medium does not contain fetal bovine serum, bovine serum albumin, or human serum albumin, as these preparations typically contain a complex mix of albumins and lipids. As used herein, the term “protein-free liquid culture medium” means a liquid culture medium that does not contain any protein (e.g., any detectable protein).

In some embodiments, a cell may be cultured for an extended period of time, e.g., for about 1 day to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, about 15 days, about 10 days, about 8 days, about 6 days, about 4 days, or about 2 days (inclusive); about 2 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, about 15 days, about 10 days, about 8 days, about 6 days, or about 4 days (inclusive); about 4 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, about 15 days, about 10 days, about 8 days, or about 6 days (inclusive); about 6 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, about 15 days, about 10 days, or about 8 days (inclusive); about 8 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, about 15 days, or about 10 days (inclusive); about 10 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, about 20 days, or about 15 days (inclusive); about 15 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, about 25 days, or about 20 days (inclusive); about 20 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, about 30 days, about 20 days, or about 25 days (inclusive); about 25 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, about 35 days, or about 30 days (inclusive); about 30 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, about 40 days, or about 35 days (inclusive); about 35 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, about 50 days, about 45 days, or about 40 days (inclusive); about 40 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, about 55 days, or about 50 days (inclusive); about 50 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, about 60 days, or about 55 days (inclusive); about 55 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, or about 60 days (inclusive); about 60 days to about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, or about 65 days (inclusive); about 65 days to about 90 days, about 85 days, about 80 days, about 75 days, or about 70 days (inclusive); about 70 days to about 90 days, about 85 days, about 80 days, or about 75 days (inclusive); about 75 days to about 90 days, about 85 days, or about 80 days (inclusive); about 80 days to about 90 days, or about 85 days (inclusive); or about 85 days to about 90 days (inclusive)) at about 20 °C to about 40 °C (e.g., about 39 °C, about 38 °C, about 37 °C, about 36 °C, or about 35 °C (inclusive); about 34 °C to about 40 °C, about 39 °C, about 38 °C, about 37 °C, about 36 °C, about 34 °C, about 32 °C, about 30 °C, about 28 °C, about 26 °C, about 24 °C, about 22 °C, about 20 °C (inclusive); about 20 °C to about 40 °C, about 20 °C to about 38 °C, about 20 °C to about 36 °C, about 20 °C to about 34 °C, about 20 °C to about 32 °C, about 20 °C to about 30 °C, about 20 °C to about 28 °C, about 20 °C to about 26 °C, about 20 °C to about 24 °C, about 20 °C to about 22 °C (inclusive); about 22 °C to about 40 °C, about 22 °C to about 38 °C, about 22 °C to about 36 °C, about 22 °C to about 34 °C, about 22 °C to about 32 °C, about 22 °C to about 30 °C, about 22 °C to about 28 °C, about 22 °C to about 26 °C, about 22 °C to about 24 °C (inclusive); about 24 °C to about 40 °C, about 24 °C to about 38 °C, about 24 °C to about 36 °C, about 24 °C to about 34 °C, about 24 °C to about 32 °C, about 24 °C to about 30 °C, about 24 °C to about 28 °C, about 24 °C to about 26 °C (inclusive); about 26 °C to about 40 °C, about 26 °C to about 38 °C, about 26 °C to about 36 °C, about 26 °C to about 34 °C, about 26 °C to about 32 °C, about 26 °C to about 30 °C, about 26 °C to about 28 °C (inclusive); about 28 °C to about 40 °C, about 28 °C to about 38 °C, about 28 °C to about 36 °C, about 28 °C to about 34 °C, about 28 °C to about 32 °C, about 28 °C to about 30 °C, about 30 °C to about 40 °C, about 30 °C to about 38 °C, about 30 °C to about 36 °C, about 30 °C to about 34 °C, about 30 °C to about 32 °C (inclusive); about 32 °C to about 40 °C, about 32 °C to about 38 °C, about 32 °C to about 36 °C, about 32 °C to about 34 °C, about 34 °C to about 40 °C, about 34 °C to about 38 °C, about 34 °C to about 36 °C, about 36 °C to about 40 °C, about 36 °C to about 38 °C, about 38 °C to about 40 °C (inclusive); about 35 °C to about 40 °C, about 39 °C, about 38 °C, or about 37 °C (inclusive); about 36 °C to about 40 °C, about 39 °C, or about 38 °C (inclusive); about 37 °C to about 40 °C, or about 39 °C (inclusive); or about 38 °C to about 40 °C (inclusive); in about 5% to about 12% (e.g., about 5% to about 10%, about 5% to about 8%, about 5% to about 7%, about 6% to about 8%, about 6% to about 10%, or about 8% to about 10%) CO ₂ . In some embodiments, the liquid cell culture is agitated for a period of time (e.g., continuously, periodically) during culturing. The term“agitation” means stirring or otherwise moving a portion of liquid culture medium in a bioreactor. This is performed in order to, e.g., increase the dissolved oxygen concentration in the liquid culture medium in a bioreactor.

Agitation can be performed using any art known method, e.g., an instrument (e.g., a shaking, rocking, or tilting platform) or propellor. Exemplary devices and methods that can be used to perform agitation of a portion of the liquid culture medium in a bioreactor are known in the art.

In some embodiments of any of the methods described herein, the liquid cell culture is a perfusion culture. The term“perfusion culture” is a term of art and refers to the culturing of cells present in a culture vessel (e.g., any of the culture vessels described herein) by periodically or continuously removing of a first liquid culture medium and at the same time or shortly thereafter adding substantially the same volume of a second liquid culture medium to the culture vessel. In some examples, there is an incremental change (e.g., increase or decrease) in the volume of the first liquid culture medium removed and added over incremental periods (e.g., an about 24-hour period, a period of between about 1 minute and about 24-hours, or a period of greater than 24 hours) during the culturing period (e.g., the culture medium refeed rate on a daily basis). The fraction of media removed and replaced each day can vary depending on the particular cells being cultured, the initial seeding density, and the cell density at a particular period of time.

In some embodiments of any of the methods described herein, the liquid cell culture is a fed-batch culture. The term“fed-batch culture” is a term of art and refers to culturing of cells present in a culture vessel by periodically or continuously adding of a second liquid culture medium to a first liquid culture medium without substantial or significant removing the first liquid culture medium or second liquid culture medium from the liquid cell culture. The second liquid culture medium can be the same as the first liquid culture medium. In some examples of fed-batch culture, the second liquid culture medium is a concentrated form of the first liquid culture medium. In some examples of fed-batch culture, the second liquid culture medium is added as a dry powder.

The liquid cell culture can be disposed in a culture vessel. Non-limiting examples of culture vessels include: a shake flask, a shake tube, a wave bag, a T- flask, a triple flask, a spinner flask, a roller bottle, a bioreactor (e.g., a perfusion bioreactor, a fed-batch bioreactor), or a multi-well plate (e.g., a 4-well cell culture plate, a 6-well cell culture plate, a 8-well cell culture plate, a 12-well cell culture plate, a 24-well cell culture plate, a 24-well cell culture plate).

In some embodiments, the culture vessel has a total interior volume of about 0.2 mL to 10,000 L, (e.g., about 0.2 mL to about 9,000 L, about 0.2 mL to about 8,000 L, about 0.2 mL to about 7,000 L, about 0.2 mL to about 6,000 L, about 0.2 mL to about 5,000 L, about 0.2 mL to about 4,000 L, about 0.2 mL to about 3,000 L, about 0.2 mL to about 2,000 L, about 0.2 mL to about 1,000 L, about 0.2 mL to about 950 L, about 0.2 mL to about 900 L, about 0.2 mL to about 850 L, about 0.2 mL to about 800 L, about 0.2 mL to about 750 mL, about 0.2 mL to about 700 L, about 0.2 mL to about 650 mL, about 0.2 mL to about 600 L, about 0.2 mL to about 550 L, about 0.2 mL to about 500 L, about 0.2 mL to about 450 L, about 0.2 mL to about 400 L, about 0.2 mL to about 300 L, about 0.2 mL to about 200 L, about 0.2 mL to about 190 L, about 0.2 mL to about 180 L, about 0.2 mL to about 170 L, about 0.2 mL to about 160 L, about 0.2 mL to about 150 L, about 0.2 mL to about 140 L, about 0.2 mL to about 130 L, about 0.2 mL to about 120 L, about 0.2 mL to about 110L, about 0.2 mL to about 100 L, about 0.2 mL to about 90 L, about 0.2 mL to about 85 L, about 0.2 mL to about 80 L, about 0.2 mL to about 75 L, about 0.2 mL to about 70 L, about 0.2 mL to about 65 L, about 0.2 mL to about 60 L, about 0.2 mL to about 55 L, about 0.2 mL to about 50 L, about 0.2 mL to about 45 L, about 0.2 mL to about 40 L, about 0.2 mL to about 35 L, about 0.2 mL to about 30 L, about 0.2 mL to about 25 L, about 0.2 mL to about 20 L, about 0.2 mL to about 15 L, about 0.2 mL to about 10 L, about 0.2 mL to about 5 L, about 0.2 mL to about 4 L, about 0.2 mL to about 4 L, about 0.2 mL to about 3 L, about 0.2 mL to about 2 L, about 0.2 mL to about 1 L; about 0.2 mL to about 900 mL, about 0.2 mL to about 800 mL, about 0.2 mL to about 700 mL, about 0.2 mL to about 600 mL, about 0.2 mL to about 500 mL, about 0.2 mL to about 400 mL, about 0.2 mL to about 300 mL, about 0.2 mL to about 250 mL, about 0.2 mL to about 100 mL, about 0.2 mL to about 90 mL, about 0.2 mL to about 80 mL, about 0.2 mL to about 70 mL, about 0.2 mL to about 60 mL, about 0.2 mL to about 50 mL, about 0.2 mL to about 40 mL, about 0.2 mL to about 30 mL, about 0.2 mL to about 25 mL, about 0.2 mL to about 20 mL, about 0.2 mL to about 15 mL, about 0.2 mL to about 10 mL, about 0.2 mL to about 5 mL, about 0.2 mL to about 4 mL, about 0.2 mL to about 3 mL, about 0.2 mL to about 2 mL, about 0.2 mL to about 1 mL; about 0.4 mL to 10,000 L, about 0.4 mL to about 9,000 L, about 0.4 mL to about 8,000 L, about 0.4 mL to about 7,000 L, about 0.4 mL to about 6,000 L, about 0.4 mL to about 5,000 L, about 0.4 mL to about 4,000 L, about 0.4 mL to about 3,000 L, about 0.4 mL to about 2,000 L, about 0.4 mL to about 1,000 L, about 0.4 mL to about 950 L, about 0.4 mL to about 900 L, about 0.4 mL to about 850 L, about 0.4 mL to about 800 L, about 0.4 mL to about 750 L, about 0.4 mL to about 700 L, about 0.4 mL to about 650 L, about 0.4 mL to about 600 L, about 0.4 mL to about 550 L, about 0.4 mL to about 500 L, about 0.4 mL to about 450 L, about 0.4 mL to about 400 L, about 0.4 mL to about 350 L, about 0.4 mL to about 300 L, about 0.4 mL to about 250 L, about 0.4 mL to about 200 L, about 0.4 mL to about 190 L, about 0.4 mL to about 180 L, about 0.4 mL to about 170 L, about 0.4 mL to about 160 L, about 0.4 mL to about 150 L, about 0.4 mL to about 140 L, about 0.4 mL to about 130 L, about 0.4 mL to about 120 L, about 0.4 mL to about 110L, about 0.4 mL to about 100 L, about 0.4 mL to about 90 L, about 0.4 mL to about 85 L, about 0.4 mL to about 80 L, about 0.4 mL to about 75 L, about 0.4 mL to about 70 L, about 0.4 mL to about 65 L, about 0.4 mL to about 60 L, about 0.4 mL to about 55 L, about 0.4 mL to about 50 L, about 0.4 mL to about 45 L, about 0.4 mL to about 40 L, about 0.4 mL to about 35 L, about 0.4 mL to about 30 L, about 0.4 mL to about 25 L, about 0.4 mL to about 20 L, about 0.4 mL to about 15 L, about 0.4 mL to about 10 L, about 0.4 mL to about 5 L, about 0.4 mL to about 4 L, about 0.4 mL to about 4 L, about 0.4 mL to about 3 L, about 0.4 mL to about 2 L, about 0.4 mL to about 1 L; about 0.4 mL to about 900 mL, about 0.4 mL to about 800 mL, about 0.4 mL to about 700 mL, about 0.4 mL to about 600 mL, about 0.4 mL to about 500 mL, about 0.4 mL to about 400 mL, about 0.4 mL to about 300 mL, about 0.4 mL to about 250 mL, about 0.4 mL to about 100 mL, about 0.4 mL to about 90 mL, about 0.4 mL to about 80 mL, about 0.4 mL to about 70 mL, about 0.4 mL to about 60 mL, about 0.4 mL to about 50 mL, about 0.4 mL to about 40 mL, about 0.4 mL to about 30 mL, about 0.4 mL to about 25 mL, about 0.4 mL to about 20 mL, about 0.4 mL to about 15 mL, about 0.4 mL to about 10 mL, about 0.4 mL to about 5 mL, about 0.4 mL to about 4 mL, about 0.4 mL to about 3 mL, about 0.4 mL to about 2 mL, about 0.4 mL to about 1 mL; about 0.6 mL to 10,000 L, about 0.6 mL to about 9,000 L, about 0.6 mL to about 8,000 L, about 0.6 mL to about 7,000 L, about 0.6 mL to about 6,000 L, about 0.6 mL to about 5,000 L, about 0.6 mL to about 4,000 L, about 0.6 mL to about 3,000 L, about 0.6 mL to about 2,000 L, about 0.6 mL to about 1,000 L, about 0.6 mL to about 950 L, about 0.6 mL to about 900 L, about 0.6 mL to about 850 L, about 0.6 mL to about 800 L, about 0.6 mL to about 750 L, about 0.6 mL to about 700 L, about 0.6 mL to about 650 L, about 0.6 mL to about 600 L, about 0.6 mL to about 550 L, about 0.6 mL to about 500 L, about 0.6 mL to about 450 L, about 0.6 mL to about 400 L, about 0.6 mL to about 350 L, about 0.6 mL to about 300 L, about 0.6 mL to about 250 L, about 0.6 mL to about 200 L, about 0.6 mL to about 190 L, about 0.6 mL to about 180 L, about 0.6 mL to about 170 L, about 0.6 mL to about 160 L, about 0.6 mL to about 150 L, about 0.6 mL to about 140 L, about 0.6 mL to about 130 L, about 0.6 mL to about 120 L, about 0.6 mL to about 110L, about 0.6 mL to about 100 L, about 0.6 mL to about 90 L, about 0.6 mL to about 85 L, about 0.6 mL to about 80 L, about 0.6 mL to about 75 L, about 0.6 mL to about 70 L, about 0.6 mL to about 65 L, about 0.6 mL to about 60 L, about 0.6 mL to about 55 L, about 0.6 mL to about 50 L, about 0.6 mL to about 45 L, about 0.6 mL to about 40 L, about 0.6 mL to about 35 L, about 0.6 mL to about 30 L, about 0.6 mL to about 25 L, about 0.6 mL to about 20 L, about 0.6 mL to about 15 L, about 0.6 mL to about 10 L, about 0.6 mL to about 5 L, about 0.6 mL to about 4 L, about 0.6 mL to about 4 L, about 0.6 mL to about 3 L, about 0.6 mL to about 2 L, about 0.6 mL to about 1 L; about 0.6 mL to about 900 mL, about 0.6 mL to about 800 mL, about 0.6 mL to about 700 mL, about 0.6 mL to about 600 mL, about 0.6 mL to about 500 mL, about 0.6 mL to about 400 mL, about 0.6 mL to about 300 mL, about 0.6 mL to about 250 mL, about 0.6 mL to about 100 mL, about 0.6 mL to about 90 mL, about 0.6 mL to about 80 mL, about 0.6 mL to about 70 mL, about 0.6 mL to about 60 mL, about 0.6 mL to about 50 mL, about 0.6 mL to about 40 mL, about 0.6 mL to about 30 mL, about 0.6 mL to about 25 mL, about 0.6 mL to about 20 mL, about 0.6 mL to about 15 mL, about 0.6 mL to about 10 mL, about 0.6 mL to about 5 mL, about 0.6 mL to about 4 mL, about 0.6 mL to about 3 mL, about 0.6 mL to about 2 mL, about 0.6 mL to about 1 mL; about 0.8 mL to 10,000 L, about 0.8 mL to about 9,000 L, about 0.8 mL to about 8,000 L, about 0.8 mL to about 7,000 L, about 0.8 mL to about 6,000 L, about 0.8 mL to about 5,000 L, about 0.8 mL to about 4,000 L, about 0.8 mL to about 3,000 L, about 0.8 mL to about 2,000 L, about 0.8 mL to about 1,000 L, about 0.5 mL to about 950 L, about 0.8 mL to about 900 L, about 0.8 mL to about 850 L, about 0.8 mL to about 800 L, about 0.8 mL to about 750 L, about 0.8 mL to about 700 L, about 0.8 mL to about 650 L, about 0.8 mL to about 600 L, about 0.8 mL to about 550 L, about 0.8 mL to about 500 L, about 0.8 mL to about 450 L, about 0.8 mL to about 400 L, about 0.8 mL to about 350 L, about 0.8 mL to about 300 L, about 0.8 mL to about 250 L, about 0.8 mL to about 200 L, about 0.8 mL to about 190 L, about 0.8 mL to about 180 L, about 0.8 mL to about 170 L, about 0.8 mL to about 160 L, about 0.8 mL to about 150 L, about 0.8 mL to about 140 L, about 0.8 mL to about 130 L, about 0.8 mL to about 120 L, about 0.8 mL to about 110L, about 0.8 mL to about 100 L, about 0.8 mL to about 90 L, about 0.8 mL to about 85 L, about 0.8 mL to about 80 L, about 0.8 mL to about 75 L, about 0.8 mL to about 70 L, about 0.8 mL to about 65 L, about 0.8 mL to about 60 L, about 0.8 mL to about 55 L, about 0.8 mL to about 50 L, about 0.8 mL to about 45 L, about 0.8 mL to about 40 L, about 0.8 mL to about 35 L, about 0.8 mL to about 30 L, about 0.8 mL to about 25 L, about 0.8 mL to about 20 L, about 0.8 mL to about 15 L, about 0.8 mL to about 10 L, about 0.8 mL to about 5 L, about 0.8 mL to about 4 L, about 0.8 mL to about 4 L, about 0.8 mL to about 3 L, about 0.8 mL to about 2 L, about 0.8 mL to about 1 L; about 0.8 mL to about 900 mL, about 0.8 mL to about 800 mL, about 0.8 mL to about 700 mL, about 0.8 mL to about 600 mL, about 0.8 mL to about 500 mL, about 0.8 mL to about 400 mL, about 0.8 mL to about 300 mL, about 0.8 mL to about 250 mL, about 0.8 mL to about 100 mL, about 0.8 mL to about 90 mL, about 0.8 mL to about 80 mL, about 0.8 mL to about 70 mL, about 0.8 mL to about 60 mL, about 0.8 mL to about 50 mL, about 0.8 mL to about 40 mL, about 0.8 mL to about 30 mL, about 0.8 mL to about 25 mL, about 0.8 mL to about 20 mL, about 0.8 mL to about 15 mL, about 0.8 mL to about 10 mL, about 0.8 mL to about 5 mL, about 0.8 mL to about 4 mL, about 0.8 mL to about 3 mL, about 0.8 mL to about 2 mL, about 0.8 mL to about 1 mL; about 1 mL to 10,000 L, about 1 mL to about 9,000 L, about 1 mL to about 8,000 L, about 1 mL to about 7,000 L, about 1 mL to about 6,000 L, about 1 mL to about 5,000 L, about 1 mL to about 4,000 L, about 1 mL to about 3,000 L, about 1 mL to about 2,000 L, about 1 mL to about 1,000 L, about 1 mL to about 950 L, about 1 mL to about 900 L, about 1 mL to about 850 L, about 1 mL to about 800 L, about 1 mL to about 750 L, about 1 mL to about 700 L, about 1 mL to about 650 L, about 1 mL to about 600 L, about 1 mL to about 550 L, about 1 mL to about 500 L, about 1 mL to about 450 L, about 1 mL to about 400 L, about 1 mL to about 350 L, about 1 mL to about 300 L, about 1 mL to about 250 L, about 1 mL to about 200 L, about 1 mL to about 190 L, about 1 mL to about 180 L, about 1 mL to about 170 L, about 1 mL to about 160 L, about 1 mL to about 150 L, about 1 mL to about 140 L, about 1 mL to about 130 L, about 1 mL to about 120 L, about 1 mL to about 110 L, about 1 mL to about 100 L, about 1 mL to about 90 L, about 1 mL to about 80 L, about 1 mL to about 70 L, about 1 mL to about 60 L, about 1 mL to about 50 L, about 1 mL to about 40 L, about 1 mL to about 30 L, about 1 mL to about 20 L, about 1 mL to about 10 L, about 1 mL to about 5 L, about 1 mL to about 4 L, about 1 mL to about 3 L, about 1 mL to about 2 L, about 1 mL to about 1 L, about 1 mL to about 900 mL, about 1 mL to about 800 mL, about 1 mL to about 700 mL, about 1 mL to about 600 mL, about 1 mL to about 500 mL, about 1 mL to about 400 mL, about 1 mL to about 300 mL, about 1 mL to about 250 mL, about 1 mL to about 100 mL, about 1 mL to about 90 mL, about 1 mL to about 80 mL, about 1 mL to about 70 mL, about 1 mL to about 60 mL, about 1 mL to about 50 mL, about 1 mL to about 40 mL, about 1 mL to about 30 mL, about 1 mL to about 25 mL, about 1 mL to about 20 mL, about 1 mL to about 15 mL, about 1 mL to about 10 mL, about 1 mL to about 5 mL; about 1 mL to about 4 mL, about 1 mL to about 3 mL, about 1 mL to about 2 mL; about 2 mL to 10,000 L, about 2 mL to about 9,000 L, about 2 mL to about 8,000 L, about 2 mL to about 7,000 L, about 2 mL to about 6,000 L, about 2 mL to about 5,000 L, about 2 mL to about 4,000 L, about 2 mL to about 3,000 L, about 2 mL to about 2,000 L, about 2 mL to about 1,000 L, about 2 mL to about 950 L, about 2 mL to about 900 L, about 2 mL to about 850 L, about 2 mL to about 800 L, about 2 mL to about 750 L, about 2 mL to about 700 L, about 2 mL to about 650 L, about 2 mL to about 600 L, about 2 mL to about 550 L, about 2 mL to about 500 L, about 2 mL to about 450 L, about 2 mL to about 400 L, about 2 mL to about 350 L, about 2 mL to about 300 L, about 2 mL to about 250 L, about 2 mL to about 200 L; about 2 mL to about 190 L, about 2 mL to about 180 L, about 2 mL to about 170 L, about 2 mL to about 160 L, about 2 mL to about 150 L, about 2 mL to about 140 L, about 2 mL to about 130 L, about 2 mL to about 120 L, about 2 mL to about 110 L, about 2 mL to about 100 L, about 2 mL to about 90 L, about 2 mL to about 80 L, about 2 mL to about 70 L, about 2 mL to about 60 L, about 2 mL to about 50 L, about 2 mL to about 40 L, about 2 mL to about 30 L, about 2 mL to about 20 L, about 2 mL to about 10 L, about 2 mL to about 5 L, about 2 mL to about 4 L, about 2 mL to about 3 L, about 2 mL to about 2 L, about 2 mL to about 1 L, about 2 mL to about 900 mL, about 2 mL to about 800 mL, about 2 mL to about 700 mL, about 2 mL to about 600 mL, about 2 mL to about 500 mL, about 2 mL to about 400 mL, about 2 mL to about 300 mL, about 2 mL to about 250 mL, about 2 mL to about 100 mL, about 2 mL to about 90 mL, about 2 mL to about 80 mL, about 2 mL to about 70 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 40 mL, about 2 mL to about 30 mL, about 2 mL to about 25 mL, about 2 mL to about 20 mL, about 2 mL to about 15 mL, about 2 mL to about 10 mL, about 2 mL to about 5 mL;

about 5 mL to 10,000 L, about 5 mL to about 9,000 L, about 5 mL to about 8,000 L, about 5 mL to about 7,000 L, about 5 mL to about 6,000 L, about 5 mL to about 5,000 L, about 5 mL to about 4,000 L, about 5 mL to about 3,000 L, about 5 mL to about 2,000 L, about 5 mL to about 1,000 L, about 5 mL to about 950 L, about 5 mL to about 900 L, about 5 mL to about 850 L, about 5 mL to about 800 L, about 5 mL to about 750 L, about 5 mL to about 700 L, about 5 mL to about 650 L, about 5 mL to about 600 L, about 5 mL to about 550 L, about 5 mL to about 500 L, about 5 mL to about 450 L, about 5 mL to about 400 L, about 5 mL to about 350 L, about 5 mL to about 300 L, about 5 mL to about 250 L, about 5 mL to about 200 L; about 5 mL to about 190 L, about 5 mL to about 180 L, about 5 mL to about 170 L, about 5 mL to about 160 L, about 5 mL to about 150 L, about 5 mL to about 140 L, about 5 mL to about 130 L, about 5 mL to about 120 L, about 5 mL to about 110 L, about 5 mL to about 100 L, about 5 mL to about 90 L, about 5 mL to about 80 L, about 5 mL to about 70 L, about 5 mL to about 60 L, about 5 mL to about 50 L, about 5 mL to about 40 L, about 5 mL to about 30 L, about 5 mL to about 20 L, about 5 mL to about 10 L, about 5 mL to about 5 L, about 5 mL to about 4 L, about 5 mL to about 3 L, about 5 mL to about 5 L, about 5 mL to about 1 L, about 5 mL to about 900 mL, about 5 mL to about 800 mL, about 5 mL to about 700 mL, about 5 mL to about 600 mL, about 5 mL to about 500 mL, about 5 mL to about 400 mL, about 5 mL to about 300 mL, about 5 mL to about 250 mL, about 5 mL to about 100 mL, about 5 mL to about 90 mL, about 5 mL to about 80 mL, about 5 mL to about 70 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 40 mL, about 5 mL to about 30 mL, about 5 mL to about 25 mL, about 5 mL to about 20 mL, about 5 mL to about 15 mL, about 5 mL to about 10 mL; about 10 mL to 10,000 L, about 10 mL to about 9,000 L, about 10 mL to about 8,000 L, about 10 mL to about 7,000 L, about 10 mL to about 6,000 L, about 10 mL to about 5,000 L, about 10 mL to about 4,000 L, about 10 mL to about 3,000 L, about 10 mL to about 2,000 L, about 10 mL to about 1,000 L, about 10 mL to about 950 L, about 10 mL to about 900 L, about 10 mL to about 850 L, about 10 mL to about 800 L, about 10 mL to about 750 L, about 10 mL to about 700 L, about 10 mL to about 650 L, about 10 mL to about 600 L, about 10 mL to about 550 L, about 10 mL to about 500 L, about 10 mL to about 450 L, about 10 mL to about 400 L, about 10 mL to about 350 L, about 10 mL to about 300 L, about 10 mL to about 250 L, about 10 mL to about 200 L, about 10 mL to about 190 L, about 10 mL to about 180 L, about 10 mL to about 170 L, about 10 mL to about 160 L, about 10 mL to about 150 L, about 10 mL to about 140 L, about 10 mL to about 130 L, about 10 mL to about 120 L, about 10 mL to about 110 L, about 10 mL to about 100 L, about 10 mL to about 90 L, about 10 mL to about 80 L, about 10 mL to about 70 L, about 10 mL to about 60 L, about 10 mL to about 50 L, about 10 mL to about 40 L, about 10 mL to about 30 L, about 10 mL to about 20 mL, about 10 mL to about 10 L, about 10 mL to about 5 L, about 10 mL to about 4 L, about 10 mL to about 3 L, about 10 mL to about 5 L, about 10 mL to about 1 L, about 10 mL to about 900 mL, about 10 mL to about 800 mL, about 10 mL to about 700 mL, about 10 mL to about 600 mL, about 10 mL to about 500 mL, about 10 mL to about 400 mL, about 10 mL to about 300 mL, about 10 mL to about 250 mL, about 10 mL to about 100 mL, about 10 mL to about 90 mL, about 10 mL to about 80 mL, about 10 mL to about 70 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 40 mL, about 10 mL to about 30 mL, about 10 mL to about 25 mL, about 10 mL to about 20 mL, about 10 mL to about 15 mL; about 25 mL to 10,000 L, about 25 mL to about 9,000 L, about 25 mL to about 8,000 L, about 25 mL to about 7,000 L, about 25 mL to about 6,000 L, about 25 mL to about 5,000 L, about 25 mL to about 4,000 L, about 25 mL to about 3,000 L, about 25 mL to about 2,000 L, about 25 mL to about 1,000 L, about 25 mL to about 950 L, about 25 mL to about 900 L, about 25 mL to about 850 L, about 25 mL to about 800 L, about 25 mL to about 750 L, about 25 mL to about 700 L, about 25 mL to about 650 L, about 25 mL to about 600 L, about 25 mL to about 550 L, about 25 mL to about 500 L, about 25 mL to about 450 L, about 25 mL to about 400 L, about 25 mL to about 350 L, about 25 mL to about 300 L, about 25 mL to about 250 L, about 25 mL to about 200 L, about 25 mL to about 190 L, about 25 mL to about 180 L, about 25 mL to about 170 L, about 25 mL to about 160 L, about 25 mL to about 150 L, about 25 mL to about 140 L, about 25 mL to about 130 L, about 25 mL to about 120 L, about 25 mL to about 110 L, about 25 mL to about 100 L, about 25 mL to about 90 L, about 25 mL to about 80 L, about 25 mL to about 70 L, about 25 mL to about 60 L, about 25 mL to about 50 L, about 25 mL to about 40 L, about 25 mL to about 30 L, about 25 mL to about 20 L, about 25 mL to about 10 L, about 25 mL to about 5 L, about 25 mL to about 4 L, about 25 mL to about 3 L, about 25 mL to about 5 L, about 25 mL to about 1 L, about 25 mL to about 900 mL, about 25 mL to about 800 mL, about 25 mL to about 700 mL, about 25 mL to about 600 mL, about 25 mL to about 500 mL, about 25 mL to about 400 mL, about 25 mL to about 300 mL, about 25 mL to about 250 mL, about 25 mL to about 100 mL, about 25 mL to about 90 mL, about 25 mL to about 80 mL, about 25 mL to about 70 mL, about 25 mL to about 60 mL, about 25 mL to about 50 mL, about 25 mL to about 40 mL, about 25 mL to about 30 mL; about 50 mL to 10,000 L, about 50 mL to about 9,000 L, about 50 mL to about 8,000 L, about 50 mL to about 7,000 L, about 50 mL to about 6,000 L, about 50 mL to about 5,000 L, about 50 mL to about 4,000 L, about 50 mL to about 3,000 L, about 50 mL to about 2,000 L, about 50 mL to about 1,000 L, about 50 mL to about 950 L, about 50 mL to about 900 L, about 50 mL to about 850 L, about 50 mL to about 800 L, about 50 mL to about 750 L, about 50 mL to about 700 L, about 50 mL to about 650 L, about 50 mL to about 600 L, about 50 mL to about 550 L, about 50 mL to about 500 L, about 50 mL to about 450 L, about 50 mL to about 400 L, about 50 mL to about 350 L, about 50 mL to about 300 L, about 50 mL to about 250 L, about 50 mL to about 200 L, about 50 mL to about 190 L, about 50 mL to about 180 L, about 50 mL to about 170 L, about 50 mL to about 160 L, about 50 mL to about 150 L, about 50 mL to about 140 L, about 50 mL to about 130 L, about 50 mL to about 120 L, about 50 mL to about 110 L, about 50 mL to about 100 L, about 50 mL to about 90 L, about 50 mL to about 80 L, about 50 mL to about 70 L, about 50 mL to about 60 L, about 50 mL to about 50 L, about 50 mL to about 40 L, about 50 mL to about 30 L, about 50 mL to about 20 L, about 50 mL to about 10 L, about 50 mL to about 5 L, about 50 mL to about 4 L, about 50 mL to about 3 L, about 50 mL to about 5 L, about 50 mL to about 1 L, about 50 mL to about 900 mL, about 50 mL to about 800 mL, about 50 mL to about 700 mL, about 50 mL to about 600 mL, about 50 mL to about 500 mL, about 50 mL to about 400 mL, about 50 mL to about 300 mL, about 50 mL to about 250 mL, about 50 mL to about 100 mL, about 50 mL to about 90 mL, about 50 mL to about 80 mL, about 50 mL to about 70 mL, about 50 mL to about 60 mL; about 75 mL to 10,000 L, about 75 mL to about 9,000 L, about 75 mL to about 8,000 L, about 75 mL to about 7,000 L, about 75 mL to about 6,000 L, about 75 mL to about 5,000 L, about 75 mL to about 4,000 L, about 75 mL to about 3,000 L, about 75 mL to about 2,000 L, about 75 mL to about 1,000 L, about 75 mL to about 950 L, about 75 mL to about 900 L, about 75 mL to about 850 L, about 75 mL to about 800 L, about 75 mL to about 750 L, about 75 mL to about 700 L, about 75 mL to about 650 L, about 75 mL to about 600 L, about 75 mL to about 550 L, about 75 mL to about 500 L, about 75 mL to about 450 L, about 75 mL to about 400 L, about 75 mL to about 350 L, about 75 mL to about 300 L, about 75 mL to about 250 L, about 75 mL to about 200 L, about 75 mL to about 190 L, about 75 mL to about 180 L, about 75 mL to about 170 L, about 75 mL to about 160 L, about 75 mL to about 150 L, about 75 mL to about 140 L, about 75 mL to about 130 L, about 75 mL to about 120 L, about 75 mL to about 110 L, about 75 mL to about 100 L, about 75 mL to about 90 L, about 75 mL to about 80 L, about 75 mL to about 70 L, about 75 mL to about 60 L, about 75 mL to about 50 L, about 75 mL to about 40 L, about 75 mL to about 30 L, about 75 mL to about 20 L, about 75 mL to about 10 L, about 75 mL to about 5 L, about 75 mL to about 4 L, about 75 mL to about 3 L, about 75 mL to about 5 L, about 75 mL to about 1 L, about 75 mL to about 900 mL, about 75 mL to about 800 mL, about 75 mL to about 700 mL, about 75 mL to about 600 mL, about 75 mL to about 500 mL, about 75 mL to about 400 mL, about 75 mL to about 300 mL, about 75 mL to about 250 mL, about 75 mL to about 100 mL, about 75 mL to about 90 mL; about 100 mL to 10,000 L, about 100 mL to about 9,000 L, about 100 mL to about 8,000 L, about 100 mL to about 7,000 L, about 100 mL to about 6,000 L, about 100 mL to about 5,000 L, about 100 mL to about 4,000 L, about 100 mL to about 3,000 L, about 100 mL to about 2,000 L, about 100 mL to about 1,000 L, about 100 mL to about 950 L, about 100 mL to about 900 L, about 100 mL to about 850 L, about 100 mL to about 800 L, about 100 mL to about 750 L, about 100 mL to about 700 L, about 100 mL to about 650 L, about 100 mL to about 600 L, about 100 mL to about 550 L, about 100 mL to about 500 L, about 100 mL to about 450 L, about 100 mL to about 400 L, about 100 mL to about 350 L, about 100 mL to about 300 L, about 100 mL to about 250 L, about 100 mL to about 200 L, about 100 mL to about 190 L, about 100 mL to about 180 L, about 100 mL to about 170 L, about 100 mL to about 160 L, about 100 mL to about 150 L, about 100 mL to about 140 L, about 100 mL to about 130 L, about 100 mL to about 120 L, about 100 mL to about 110 L, about 100 mL to about 100 L, about 100 mL to about 90 L, about 100 mL to about 80 L, about 100 mL to about 70 L, about 100 mL to about 60 L, about 100 mL to about 50 L, about 100 mL to about 40 L, about 100 mL to about 30 L, about 100 mL to about 20 L, about 100 mL to about 10 L, about 100 mL to about 5 L, about 100 mL to about 4 L, about 100 mL to about 3 L, about 100 mL to about 5 L, about 100 mL to about 1 L, about 100 mL to about 900 mL, about 100 mL to about 800 mL, about 100 mL to about 700 mL, about 100 mL to about 600 mL, about 100 mL to about 500 mL, about 100 mL to about 400 mL, about 100 mL to about 300 mL, about 100 mL to about 250 mL; about 250 mL to 10,000 L, about 250 mL to about 9,000 L, about 250 mL to about 8,000 L, about 250 mL to about 7,000 L, about 250 mL to about 6,000 L, about 250 mL to about 5,000 L, about 250 mL to about 4,000 L, about 250 mL to about 3,000 L, about 250 mL to about 2,000 L, about 250 mL to about 1,000 L, about 250 mL to about 950 L, about 250 mL to about 900 L, about 250 mL to about 850 L, about 250 mL to about 800 L, about 250 mL to about 750 L, about 250 mL to about 700 L, about 250 mL to about 650 L, about 250 mL to about 600 L, about 250 mL to about 550 L, about 250 mL to about 500 L, about 250 mL to about 450 L, about 250 mL to about 400 L, about 250 mL to about 350 L, about 250 mL to about 300 L, about 250 mL to about 250 L, about 250 mL to about 200 L, about 250 mL to about 190 L, about 250 mL to about 180 L, about 250 mL to about 170 L, about 250 mL to about 160 L, about 250 mL to about 150 L, about 250 mL to about 140 L, about 250 mL to about 130 L, about 250 mL to about 120 L, about 250 mL to about 110 L, about 250 mL to about 100 L, about 250 mL to about 90 L, about 250 mL to about 80 L, about 250 mL to about 70 L, about 250 mL to about 60 L, about 250 mL to about 50 L, about 250 mL to about 40 L, about 250 mL to about 30 L, about 250 mL to about 20 L, about 250 mL to about 10 L, about 250 mL to about 5 L, about 250 mL to about 4 L, about 250 mL to about 3 L, about 250 mL to about 5 L, about 250 mL to about 1 L, about 250 mL to about 900 mL, about 250 mL to about 800 mL, about 250 mL to about 700 mL, about 250 mL to about 600 mL, about 250 mL to about 500 mL, about 250 mL to about 400 mL, about 250 mL to about 300 mL; about 300 mL to 10,000 L, about 300 mL to about 9,000 L, about 300 mL to about 8,000 L, about 300 mL to about 7,000 L, about 300 mL to about 6,000 L, about 300 mL to about 5,000 L, about 300 mL to about 4,000 L, about 300 mL to about 3,000 L, about 300 mL to about 2,000 L, about 300 mL to about 1,000 L, about 300 mL to about 950 L, about 300 mL to about 900 L, about 300 mL to about 850 L, about 300 mL to about 800 L, about 300 mL to about 750 L, about 300 mL to about 700 L, about 300 mL to about 650 L, about 300 mL to about 600 L, about 300 mL to about 550 L, about 300 mL to about 500 L, about 300 mL to about 450 L, about 300 mL to about 400 L, about 300 mL to about 350 L, about 300 mL to about 300 L, about 300 mL to about 250 L, about 300 mL to about 200 L, about 300 mL to about 190 L, about 300 mL to about 180 L, about 300 mL to about 170 L, about 300 mL to about 160 L, about 300 mL to about 150 L, about 300 mL to about 140 L, about 300 mL to about 130 L, about 300 mL to about 120 L, about 300 mL to about 110 L, about 300 mL to about 100 L, about 300 mL to about 90 L, about 300 mL to about 80 L, about 300 mL to about 70 L, about 300 mL to about 60 L, about 300 mL to about 50 L, about 300 mL to about 40 L, about 300 mL to about 30 L, about 300 mL to about 20 L, about 300 mL to about 10 L, about 300 mL to about 5 L, about 300 mL to about 4 L, about 300 mL to about 3 L, about 300 mL to about 5 L, about 300 mL to about 1 L, about 300 mL to about 900 mL, about 300 mL to about 800 mL, about 300 mL to about 700 mL, about 300 mL to about 600 mL, about 300 mL to about 500 mL, about 300 mL to about 400 mL; about 450 mL to 10,000 L, about 450 mL to about 9,000 L, about 450 mL to about 8,000 L, about 450 mL to about 7,000 L, about 450 mL to about 6,000 L, about 450 mL to about 5,000 L, about 450 mL to about 4,000 L, about 450 mL to about 3,000 L, about 450 mL to about 2,000 L, about 450 mL to about 1,000 L, about 450 mL to about 950 L, about 450 mL to about 900 L, about 450 mL to about 850 L, about 450 mL to about 800 L, about 450 mL to about 750 L, about 450 mL to about 700 L, about 450 mL to about 650 L, about 450 mL to about 600 L, about 450 mL to about 550 L, about 450 mL to about 500 L, about 450 mL to about 450 L, about 450 mL to about 400 L, about 450 mL to about 350 L, about 450 mL to about 300 L, about 450 mL to about 250 L, about 450 mL to about 200 L, about 450 mL to about 190 L, about 450 mL to about 180 L, about 450 mL to about 170 L, about 450 mL to about 160 L, about 450 mL to about 150 L, about 450 mL to about 140 L, about 450 mL to about 130 L, about 450 mL to about 120 L, about 450 mL to about 110 L, about 450 mL to about 100 L, about 450 mL to about 90 L, about 450 mL to about 80 L, about 450 mL to about 70 L, about 450 mL to about 60 L, about 450 mL to about 50 L, about 450 mL to about 40 L, about 450 mL to about 30 L, about 450 mL to about 20 L, about 450 mL to about 10 L, about 450 mL to about 5 L, about 450 mL to about 4 L, about 450 mL to about 3 L, about 450 mL to about 5 L, about 450 mL to about 1 L, about 450 mL to about 900 mL, about 450 mL to about 800 mL, about 450 mL to about 700 mL, about 450 mL to about 600 mL; about 500 mL to 10,000 L, about 500 mL to about 9,000 L, about 500 mL to about 8,000 L, about 500 mL to about 7,000 L, about 500 mL to about 6,000 L, about 500 mL to about 5,000 L, about 500 mL to about 4,000 L, about 500 mL to about 3,000 L, about 500 mL to about 2,000 L, about 500 mL to about 1,000 L, about 500 mL to about 950 L, about 500 mL to about 900 L, about 500 mL to about 850 L, about 500 mL to about 800 L, about 500 mL to about 750 L, about 500 mL to about 700 L, about 500 mL to about 650 L, about 500 mL to about 600 L, about 500 mL to about 550 L, about 500 mL to about 500 L, about 500 mL to about 450 L, about 500 mL to about 400 L, about 500 mL to about 350 L, about 500 mL to about 300 L, about 500 mL to about 250 L, about 500 mL to about 200 L, about 500 mL to about 190 L, about 500 mL to about 180 L, about 500 mL to about 170 L, about 500 mL to about 160 L, about 500 mL to about 150 L, about 500 mL to about 140 L, about 500 mL to about 130 L, about 500 mL to about 120 L, about 500 mL to about 110 L, about 500 mL to about 100 L, about 500 mL to about 90 L, about 500 mL to about 80 L, about 500 mL to about 70 L, about 500 mL to about 60 L, about 500 mL to about 50 L, about 500 mL to about 40 L, about 500 mL to about 30 L, about 500 mL to about 20 L, about 500 mL to about 10 L, about 500 mL to about 5 L, about 500 mL to about 4 L, about 500 mL to about 3 L, about 500 mL to about 5 L, about 500 mL to about 1 L, about 500 mL to about 900 mL, about 500 mL to about 800 mL, about 500 mL to about 700 mL, about 500 mL to about 600 mL; about 750 mL to 10,000 L, about 750 mL to about 9,000 L, about 750 mL to about 8,000 L, about 750 mL to about 7,000 L, about 750 mL to about 6,000 L, about 750 mL to about 5,000 L, about 750 mL to about 4,000 L, about 750 mL to about 3,000 L, about 750 mL to about 2,000 L, about 750 mL to about 1,000 L, about 750 mL to about 950 L, about 750 mL to about 900 L, about 750 mL to about 850 L, about 750 mL to about 800 L, about 750 mL to about 750 L, about 750 mL to about 700 L, about 750 mL to about 650 L, about 750 mL to about 600 L, about 750 mL to about 550 L, about 750 mL to about 500 L, about 750 mL to about 450 L, about 750 mL to about 400 L, about 750 mL to about 350 L, about 750 mL to about 300 L, about 750 mL to about 250 L, about 750 mL to about 200 L, about 750 mL to about 190 L, about 750 mL to about 180 L, about 750 mL to about 170 L, about 750 mL to about 160 L, about 750 mL to about 150 L, about 750 mL to about 140 L, about 750 mL to about 130 L, about 750 mL to about 120 L, about 750 mL to about 110 L, about 750 mL to about 100 L, about 750 mL to about 90 L, about 750 mL to about 80 L, about 750 mL to about 70 L, about 750 mL to about 60 L, about 750 mL to about 50 L, about 750 mL to about 40 L, about 750 mL to about 30 L, about 750 mL to about 20 L, about 750 mL to about 10 L, about 750 mL to about 5 L, about 750 mL to about 4 L, about 750 mL to about 3 L, about 750 mL to about 5 L, about 750 mL to about 1 L, about 750 mL to about 900 mL, about 750 mL to about 800 mL; about 1 L to 10,000 L, about 1 L to about 9,000 L, about 1 L to about 8,000 L, about 1 L to about 7,000 L, about 1 L to about 6,000 L, about 1 L to about 5,000 L, about 1 L to about 4,000 L, about 1 L to about 3,000 L, about 1 L to about 2,000 L, about 1 L to about 1,000 L, about 1 L to about 950 L, about 1 L to about 900 L, about 1 L to about 850 L, about 1 L to about 800 L, about 1 L to about 750 L, about 1 L to about 700 L, about 1 L to about 650 L, about 1 L to about 600 L, about 1 L to about 550 L, about 1 L to about 500 L, about 1 L to about 450 L, about 1 L to about 400 L, about 1 L to about 350 L, about 1 L to about 300 L, about 1 L to about 250 L, about 1 L to about 200 L, about 1 L to about 190 L, about 1 L to about 180 L, about 1 L to about 170 L, about 1 L to about 160 L, about 1 L to about 150 L, about 1 L to about 140 L, about 1 L to about 130 L, about 1 L to about 120 L, about 1 L to about 110 L, about 1 L to about 100 L, about 1 L to about 90 L, about 1 L to about 80 L, about 1 L to about 70 L, about 1 L to about 60 L, about 1 L to about 50 L, about 1 L to about 40 L, about 1 L to about 30 L, about 1 L to about 20 L, about 1 L to about 10 L, about 1 L to about 5 L, about 1 L to about 4 L, about 1 L to about 3 L, about 1 L to about 2L; about 2 L to 10,000 L, about 2 L to about 9,000 L, about 2 L to about 8,000 L, about 2 L to about 7,000 L, about 2 L to about 6,000 L, about 2 L to about 5,000 L, about 2 L to about 4,000 L, about 2 L to about 3,000 L, about 2 L to about 2,000 L, about 2 L to about 1,000 L, about 2 L to about 950 L, about 2 L to about 900 L, about 2 L to about 850 L, about 2 L to about 800 L, about 2 L to about 750 L, about 2 L to about 700 L, about 2 L to about 650 L, about 2 L to about 600 L, about 2 L to about 550 L, about 2 L to about 500 L, about 2 L to about 450 L, about 2 L to about 400 L, about 2 L to about 350 L, about 2 L to about 300 L, about 2 L to about 250 L, about 2 L to about 200 L, about 2 L to about 190 L, about 2 L to about 180 L, about 2 L to about 170 L, about 2 L to about 160 L, about 2 L to about 150 L, about 2 L to about 140 L, about 2 L to about 130 L, about 2 L to about 120 L, about 2 L to about 110 L, about 2 L to about 100 L, about 2 L to about 90 L, about 2 L to about 80 L, about 2 L to about 70 L, about 2 L to about 60 L, about 2 L to about 50 L, about 2 L to about 40 L, about 2 L to about 30 L, about 2 L to about 20 L, about 2 L to about 10 L, about 2 L to about 5 L, about 2 L to about 4 L, about 2 L to about 3 L; about 4 L to 10,000 L, about 4 L to about 9,000 L, about 4 L to about 8,000 L, about 4 L to about 7,000 L, about 4 L to about 6,000 L, about 4 L to about 5,000 L, about 4 L to about 4,000 L, about 4 L to about 3,000 L, about 4 L to about 2,000 L, about 4 L to about 1,000 L, about 4 L to about 950 L, about 4 L to about 900 L, about 4 L to about 850 L, about 4 L to about 800 L, about 4 L to about 750 L, about 4 L to about 700 L, about 4 L to about 650 L, about 4 L to about 600 L, about 4 L to about 550 L, about 4 L to about 500 L, about 4 L to about 450 L, about 4 L to about 400 L, about 4 L to about 350 L, about 4 L to about 300 L, about 4 L to about 250 L, about 4 L to about 200 L, about 4 L to about 190 L, about 4 L to about 180 L, about 4 L to about 170 L, about 4 L to about 160 L, about 4 L to about 150 L, about 4 L to about 140 L, about 4 L to about 130 L, about 4 L to about 120 L, about 4 L to about 110 L, about 4 L to about 100 L, about 4 L to about 90 L, about 4 L to about 80 L, about 4 L to about 70 L, about 4 L to about 60 L, about 4 L to about 50 L, about 4 L to about 40 L, about 4 L to about 30 L, about 4 L to about 20 L, about 4 L to about 10 L, about 4 L to about 5 L; about 5 L to 10,000 L, about 5 L to about 9,000 L, about 5 L to about 8,000 L, about 5 L to about 7,000 L, about 5 L to about 6,000 L, about 5 L to about 5,000 L, about 5 L to about 4,000 L, about 5 L to about 3,000 L, about 5 L to about 2,000 L, about 5 L to about 1,000 L, about 5 L to about 950 L, about 5 L to about 900 L, about 5 L to about 850 L, about 5 L to about 800 L, about 5 L to about 750 L, about 5 L to about 700 L, about 5 L to about 650 L, about 5 L to about 600 L, about 5 L to about 550 L, about 5 L to about 500 L, about 5 L to about 450 L, about 5 L to about 400 L, about 5 L to about 350 L, about 5 L to about 300 L, about 5 L to about 250 L, about 5 L to about 200 L, about 5 L to about 190 L, about 5 L to about 180 L, about 5 L to about 170 L, about 5 L to about 160 L, about 5 L to about 150 L, about 5 L to about 140 L, about 5 L to about 130 L, about 5 L to about 120 L, about 5 L to about 110 L, about 5 L to about 100 L, about 5 L to about 90 L, about 5 L to about 80 L, about 5 L to about 70 L, about 5 L to about 60 L, about 5 L to about 50 L, about 5 L to about 40 L, about 5 L to about 30 L, about 5 L to about 20 L, about 5 L to about 10 L, about 6 L to about 10 L, about 6 L to about 8 L, about 8 L to about 10 L; about 7.5 L to 10,000 L, about 7.5 L to about 9,000 L, about 7.5 L to about 8,000 L, about 7.5 L to about 7,000 L, about 7.5 L to about 6,000 L, about 7.5 L to about 5,000 L, about 7.5 L to about 4,000 L, about 7.5 L to about 3,000 L, about 7.5 L to about 2,000 L, about 7.5 L to about 1,000 L, about 7.5 L to about 950 L, about 7.5 L to about 900 L, about 7.5 L to about 850 L, about 7.5 L to about 800 L, about 7.5 L to about 750 L, about 7.5 L to about 700 L, about 7.5 L to about 650 L, about 7.5 L to about 600 L, about 7.5 L to about 550 L, about 7.5 L to about 500 L, about 7.5 L to about 450 L, about 7.5 L to about 400 L, about 7.5 L to about 350 L, about 7.5 L to about 300 L, about 7.5 L to about 250 L, about 7.5 L to about 200 L, about 7.5 L to about 190 L, about 7.5 L to about 180 L, about 7.5 L to about 170 L, about 7.5 L to about 160 L, about 7.5 L to about 150 L, about 7.5 L to about 140 L, about 7.5 L to about 130 L, about 7.5 L to about 120 L, about 7.5 L to about 110 L, about 7.5 L to about 100 L, about 7.5 L to about 90 L, about 7.5 L to about 80 L, about 7.5 L to about 70 L, about 7.5 L to about 60 L, about 7.5 L to about 50 L, about 7.5 L to about 40 L, about 7.5 L to about 30 L, about 7.5 L to about 20 L, about 7.5 L to about 10 L; about 10 L to 10,000 L, about 10 L to about 9,000 L, about 10 L to about 8,000 L, about 10 L to about 7,000 L, about 10 L to about 6,000 L, about 10 L to about 5,000 L, about 10 L to about 4,000 L, about 10 L to about 3,000 L, about 10 L to about 2,000 L, about 10 L to about 1,000 L, about 10 L to about 950 L, about 10 L to about 900 L, about 10 L to about 850 L, about 10 L to about 800 L, about 10 L to about 750 L, about 10 L to about 700 L, about 10 L to about 650 L, about 10 L to about 600 L, about 10 L to about 550 L, about 10 L to about 500 L, about 10 L to about 450 L, about 10 L to about 400 L, about 10 L to about 350 L, about 10 L to about 300 L, about 10 L to about 250 L, about 10 L to about 200 L, about 10 L to about 190 L, about 10 L to about 180 L, about 10 L to about 170 L, about 10 L to about 160 L, about 10 L to about 150 L, about 10 L to about 140 L, about 10 L to about 130 L, about 10 L to about 120 L, about 10 L to about 110 L, about 10 L to about 100 L, about 10 L to about 90 L, about 10 L to about 80 L, about 10 L to about 70 L, about 10 L to about 60 L, about 10 L to about 50 L, about 10 L to about 40 L, about 10 L to about 30 L, about 10 L to about 20 L, about 10 L to about 15 L, about 15 L to about 20 L, about 15 L to about 25 L, about 15 L to about 30 L; about 15 L to about 40 L, about 15 L to about 50 L, about 15 L to about 60 L, about 15 L to about 70 L, about 15 L to about 80 L, about 15 L to about 90 L, about 15 L to about 100 L, about 15 L to about 110 L, about 15 L to about 120 L, about 15 L to about 130 L, about 15 L to about 140 L, about 15 L to about 150 L, about 15 L to about 160 L, about 15 L to about 170 L, about 15 L to about 180 L, about 15 L to about 190 L; about 15 L to about 200 L, about 15 L to about 250 L, about 15 L to about 300 L, about 15 L to about 350 L, about 15 L to about 400 L, about 15 L to about 450 L, about 15 L to about 500 L, about 15 L to about 550 L, about 15 L to about 600 L, about 15 L to about 650 L, about 15 L to about 700 L, about 15 L to about 750 L, about 15 L to about 800 L, about 15 L to about 850 L, about 15 L to about 900 L, about 15 L to about 950 L, about 15 L to about 1,000 L, about 15 L to about 2,000 L, about 15 L to about 3,000 L, about 15 L to about 4,000 L, about 15 L to about 5,000 L, about 15 L to about 6,000 L, about 15 L to about 7,000 L, about 15 L to about 8,000 L, about 15 L to about 9,000 L, about 15 L to 10,000 L; about 20 L to about 25 L, about 20 L to about 30 L, about 20 L to about 40 L, about 20 L to about 50 L, about 20 L to about 60 L, about 20 L to about 70 L, about 20 L to about 80 L, about 20 L to about 90 L, about 20 L to about 100 L, about 20 L to about 110 L, about 20 L to about 120 L, about 20 L to about 130 L, about 20 L to about 140 L, about 20 L to about 150 L, about 20 L to about 160 L, about 20 L to about 170 L, about 20 L to about 180 L, about 20 L to about 190 L, about 20 L to about 200 L, about 20 L to about 250 L, about 20 L to about 300 L, about 20 L to about 350 L, about 20 L to about 400 L, about 20 L to about 450 L, about 20 L to about 500 L, about 20 L to about 550 L, about 20 L to about 600 L, about 20 L to about 650 L, about 20 L to about 700 L, about 20 L to about 750 L, about 20 L to about 800 L, about 20 L to about 850 L, about 20 L to about 900 L, about 20 L to about 950 L, about 20 L to about 1,000 L, about 20 L to about 2,000 L, about 20 L to about 3,000 L, about 20 L to about 4,000 L, about 20 L to about 5,000 L, about 20 L to about 6,000 L, about 20 L to about 7,000 L, about 20 L to about 8,000 L, about 20 L to about 9,000 L, about 20 L to 10,000 L; about 25 L to about 30 L; about 25 L to about 40 L, about 25 L to about 50 L, about 25 L to about 60 L, about 25 L to about 70 L, about 25 L to about 80 L, about 25 L to about 90 L, about 25 L to about 100 L, about 25 L to about 110 L, about 25 L to about 120 L, about 25 L to about 130 L, about 25 L to about 140 L, about 25 L to about 150 L, about 25 L to about 160 L, about 25 L to about 170 L, about 25 L to about 180 L, about 25 L to about 190 L, about 25 L to about 200 L, about 25 L to about 250 L, about 25 L to about 300 L, about 25 L to about 350 L, about 25 L to about 400 L, about 25 L to about 450 L, about 25 L to about 500 L, about 25 L to about 550 L, about 25 L to about 600 L, about 25 L to about 650 L, about 25 L to about 700 L, about 25 L to about 750 L, about 25 L to about 800 L, about 25 L to about 850 L, about 25 L to about 900 L, about 25 L to about 950 L, about 25 L to about 1,000 L, about 25 L to about 2,000 L, about 25 L to about 3,000 L, about 25 L to about 4,000 L, about 25 L to about 5,000 L, about 25 L to about 6,000 L, about 25 L to about 7,000 L, about 25 L to about 8,000 L, about 25 L to about 9,000 L, about 25 L to 10,000 L; about 30 L to about 50 L, about 30 L to about 75 L, about 30 L to about 100 L, about 30 L to about 150 L, about 30 L to about 200 L,

about 30 L to about 250 L, about 30 L to about 300 L, about 30 L to about 350 L, about 30 L to about 400 L, about 30 L to about 450 L, about 30 L to about 500 L, about 30 L to about 550 L, about 30 L to about 600 L, about 30 L to about 650 L, about 30 L to about 700 L, about 30 L to about 750 L, about 30 L to about 800 L, about 30 L to about 850 L, about 30 L to about 900 L, about 30 L to about 950 L, about 30 L to about 1,000 L, about 30 L to about 2,000 L, about 30 L to about 3,000 L, about 30 L to about 4,000 L, about 30 L to about 5,000 L, about 30 L to about 6,000 L, about 30 L to about 7,000 L, about 30 L to about 8,000 L, about 30 L to about 9,000 L, about 30 L to 10,000 L; about 40 L to about 75 L, about 40 L to about 100 L, about 40 L to about 150 L, about 40 L to about 200 L; about 40 L to about 250 L, about 40 L to about 300 L, about 40 L to about 350 L, about 40 L to about 400 L, about 40 L to about 450 L, about 40 L to about 500 L, about 40 L to about 550 L, about 40 L to about 600 L, about 40 L to about 650 L, about 40 L to about 700 L, about 40 L to about 750 L, about 40 L to about 800 L, about 40 L to about 850 L, about 40 L to about 900 L, about 40 L to about 950 L, about 40 L to about 1,000 L, about 40 L to about 2,000 L, about 40 L to about 3,000 L, about 40 L to about 4,000 L, about 40 L to about 5,000 L, about 40 L to about 6,000 L, about 40 L to about 7,000 L, about 40 L to about 8,000 L, about 40 L to about 9,000 L, about 40 L to 10,000 L; about 50 L to about 60 L, about 50 L to about 70 L, about 50 L to about 80 L, about 50 L to about 90 L, about 50 L to about 100 L, about 50 L to about 110 L, about 50 L to about 120 L, about 50 L to about 130 L, about 50 L to about 140 L, about 50 L to about 150 L, about 50 L to about 160 L, about 50 L to about 170 L, about 50 L to about 180 L, about 50 L to about 190 L, about 50 L to about 200 L, about 50 L to about 250 L, about 50 L to about 300 L, about 50 L to about 350 L, about 50 L to about 400 L, about 50 L to about 450 L, about 50 L to about 500 L, about 50 L to about 550 L, about 50 L to about 600 L, about 50 L to about 650 L, about 50 L to about 700 L, about 50 L to about 750 L, about 50 L to about 800 L, about 50 L to about 850 L, about 50 L to about 900 L, about 50 L to about 950 L, about 50 L to about 1,000 L, about 50 L to about 2,000 L, about 50 L to about 3,000 L, about 50 L to about 4,000 L, about 50 L to about 5,000 L, about 50 L to about 6,000 L, about 50 L to about 7,000 L, about 50 L to about 8,000 L, about 50 L to about 9,000 L, about 50 L to 10,000 L; about 75 L to about 100 L, about 75 L to about 110 L, about 75 L to about 120 L, about 75 L to about 130 L, about 75 L to about 140 L, about 75 L to about 150 L, about 75 L to about 160 L, about 75 L to about 170 L, about 75 L to about 180 L, about 75 L to about 190 L, about 75 L to about 200 L, about 75 L to about 250 L, about 75 L to about 300 L, about 75 L to about 350 L, about 75 L to about 400 L, about 75 L to about 450 L, about 75 L to about 500 L, about 75 L to about 550 L, about 75 L to about 600 L, about 75 L to about 650 L, about 75 L to about 700 L, about 75 L to about 750 L, about 75 L to about 800 L, about 75 L to about 850 L, about 75 L to about 900 L, about 75 L to about 950 L, about 75 L to about 1,000 L, about 75 L to about 2,000 L, about 75 L to about 3,000 L, about 75 L to about 4,000 L, about 75 L to about 5,000 L, about 75 L to about 6,000 L, about 75 L to about 7,000 L, about 75 L to about 8,000 L, about 75 L to about 9,000 L, about 75 L to 10,000 L; about 100 L to about 110 L, about 100 L to about 120 L, about 100 L to about 130 L, about 100 L to about 140 L, about 100 L to about 150 L, about 100 L to about 160 L, about 100 L to about 170 L, about 100 L to about 180 L, about 100 L to about 190 L, about 100 L to about 200 L, about 100 L to about 250 L, about 100 L to about 300 L, about 100 L to about 350 L, about 100 L to about 400 L, about 100 L to about 450 L, about 100 L to about 500 L, about 100 L to about 550 L, about 100 L to about 600 L, about 100 L to about 650 L, about 100 L to about 700 L, about 100 L to about 750 L, about 100 L to about 800 L, about 100 L to about 850 L, about 100 L to about 900 L, about 100 L to about 950 L, about 100 L to about 1,000 L, about 100 L to about 2,000 L, about 100 L to about 3,000 L, about 100 L to about 4,000 L, about 100 L to about 5,000 L, about 100 L to about 6,000 L, about 100 L to about 7,000 L, about 100 L to about 8,000 L, about 100 L to about 9,000 L, about 100 L to 10,000 L; about 120 L to about 130 L, about 120 L to about 140 L, about 120 L to about 150 L, about 120 L to about 160 L, about 120 L to about 170 L, about 120 L to about 180 L, about 120 L to about 190 L, about 120 L to about 200 L, about 120 L to about 250 L, about 120 L to about 300 L, about 120 L to about 350 L, about 120 L to about 400 L, about 120 L to about 450 L, about 120 L to about 500 L, about 120 L to about 550 L, about 120 L to about 600 L, about 120 L to about 650 L, about 120 L to about 700 L, about 120 L to about 750 L, about 120 L to about 800 L, about 120 L to about 850 L, about 120 L to about 900 L, about 120 L to about 950 L, about 120 L to about 1,000 L, about 120 L to about 2,000 L, about 120 L to about 3,000 L, about 120 L to about 4,000 L, about 120 L to about 5,000 L, about 120 L to about 6,000 L, about 120 L to about 7,000 L, about 120 L to about 8,000 L, about 120 L to about 9,000 L, about 120 L to 10,000 L; about 150 L to about 160 L, about 150 L to about 170 L, about 150 L to about 180 L, about 150 L to about 190 L, about 150 L to about 200 L, about 150 L to about 250 L, about 150 L to about 300 L, about 150 L to about 350 L, about 150 L to about 400 L, about 150 L to about 450 L, about 150 L to about 500 L, about 150 L to about 550 L, about 150 L to about 600 L, about 150 L to about 650 L, about 150 L to about 700 L, about 150 L to about 750 L, about 150 L to about 800 L, about 150 L to about 850 L, about 150 L to about 900 L, about 150 L to about 950 L, about 150 L to about 1,000 L, about 150 L to about 2,000 L, about 150 L to about 3,000 L, about 150 L to about 4,000 L, about 150 L to about 5,000 L, about 150 L to about 6,000 L, about 150 L to about 7,000 L, about 150 L to about 8,000 L, about 150 L to about 9,000 L, about 150 L to 10,000 L; about 200 L to about 250 L, about 200 L to about 300 L, about 200 L to about 350 L, about 200 L to about 400 L, about 200 L to about 450 L, about 200 L to about 500 L, about 200 L to about 550 L, about 200 L to about 600 L, about 200 L to about 650 L, about 200 L to about 700 L, about 200 L to about 750 L, about 200 L to about 800 L, about 200 L to about 850 L, about 200 L to about 900 L, about 200 L to about 950 L, about 200 L to about 1,000 L, about 200 L to about 2,000 L, about 200 L to about 3,000 L, about 200 L to about 4,000 L, about 200 L to about 5,000 L, about 200 L to about 6,000 L, about 200 L to about 7,000 L, about 200 L to about 8,000 L, about 200 L to about 9,000 L, about 200 L to 10,000 L; about 250 L to about 300 L, about 250 L to about 350 L, about 250 L to about 400 L, about 250 L to about 450 L, about 250 L to about 500 L, about 250 L to about 550 L, about 250 L to about 600 L, about 250 L to about 650 L, about 250 L to about 700 L, about 250 L to about 750 L, about 250 L to about 800 L, about 250 L to about 850 L, about 250 L to about 900 L, about 250 L to about 950 L, about 250 L to about 1,000 L, about 250 L to about 2,000 L, about 250 L to about 3,000 L, about 250 L to about 4,000 L, about 250 L to about 5,000 L, about 250 L to about 6,000 L, about 250 L to about 7,000 L, about 250 L to about 8,000 L, about 250 L to about 9,000 L, about 250 L to 10,000 L; about 300 L to about 350 L, about 300 L to about 400 L, about 300 L to about 450 L, about 300 L to about 500 L, about 300 L to about 550 L, about 300 L to about 600 L, about 300 L to about 650 L, about 300 L to about 700 L, about 300 L to about 750 L, about 300 L to about 800 L, about 300 L to about 850 L, about 300 L to about 900 L, about 300 L to about 950 L, about 300 L to about 1,000 L, about 300 L to about 2,000 L, about 300 L to about 3,000 L, about 300 L to about 4,000 L, about 300 L to about 5,000 L, about 300 L to about 6,000 L, about 300 L to about 7,000 L, about 300 L to about 8,000 L, about 300 L to about 9,000 L, about 300 L to 10,000 L; about 350 L to about 400 L, about 350 L to about 450 L, about 350 L to about 500 L, about 350 L to about 550 L, about 350 L to about 600 L, about 350 L to about 650 L, about 350 L to about 700 L, about 350 L to about 750 L, about 350 L to about 800 L, about 350 L to about 850 L, about 350 L to about 900 L, about 350 L to about 950 L, about 350 L to about 1,000 L, about 350 L to about 2,000 L, about 350 L to about 3,000 L, about 350 L to about 4,000 L, about 350 L to about 5,000 L, about 350 L to about 6,000 L, about 350 L to about 7,000 L, about 350 L to about 8,000 L, about 350 L to about 9,000 L, about 350 L to 10,000 L; about 400 L to about 450 L, about 400 L to about 500 L, about 400 L to about 550 L, about 400 L to about 600 L, about 400 L to about 650 L, about 400 L to about 700 L, about 400 L to about 750 L, about 400 L to about 800 L, about 400 L to about 850 L, about 400 L to about 900 L, about 400 L to about 950 L, about 400 L to about 1,000 L, about 400 L to about 2,000 L, about 400 L to about 3,000 L, about 400 L to about 4,000 L, about 400 L to about 5,000 L, about 400 L to about 6,000 L, about 400 L to about 7,000 L, about 400 L to about 8,000 L, about 400 L to about 9,000 L, about 400 L to 10,000 L; about 450 L to about 500 L, about 450 L to about 550 L, about 450 L to about 600 L, about 450 L to about 650 L, about 450 L to about 700 L, about 450 L to about 750 L, about 450 L to about 800 L, about 450 L to about 850 L, about 450 L to about 900 L, about 450 L to about 950 L, about 450 L to about 1,000 L, about 450 L to about 2,000 L, about 450 L to about 3,000 L, about 450 L to about 4,000 L, about 450 L to about 5,000 L, about 450 L to about 6,000 L, about 450 L to about 7,000 L, about 450 L to about 8,000 L, about 450 L to about 9,000 L, about 450 L to 10,000 L; about 500 L to about 550 L, about 500 L to about 600 L, about 500 L to about 650 L, about 500 L to about 700 L, about 500 L to about 750 L, about 500 L to about 800 L, about 500 L to about 850 L, about 500 L to about 900 L, about 500 L to about 950 L, about 500 L to about 1,000 L, about 500 L to about 2,000 L, about 500 L to about 3,000 L, about 500 L to about 4,000 L, about 500 L to about 5,000 L, about 500 L to about 6,000 L, about 500 L to about 7,000 L, about 500 L to about 8,000 L, about 500 L to about 9,000 L, about 500 L to 10,000 L; about 600 L to about 650 L, about 600 L to about 700 L, about 600 L to about 750 L, about 600 L to about 800 L, about 600 L to about 850 L, about 600 L to about 900 L, about 600 L to about 950 L, about 600 L to about 1,000 L, about 600 L to about 2,000 L, about 600 L to about 3,000 L, about 600 L to about 4,000 L, about 600 L to about 5,000 L, about 600 L to about 6,000 L, about 600 L to about 7,000 L, about 600 L to about 8,000 L, about 600 L to about 9,000 L, about 600 L to 10,000 L; about 700 L to about 750 L, about 700 L to about 800 L, about 700 L to about 850 L, about 700 L to about 900 L, about 700 L to about 950 L, about 700 L to about 1,000 L, about 700 L to about 2,000 L, about 700 L to about 3,000 L, about 700 L to about 4,000 L, about 700 L to about 5,000 L, about 700 L to about 6,000 L, about 700 L to about 7,000 L, about 700 L to about 8,000 L, about 700 L to about 9,000 L, about 700 L to 10,000 L; about 800 L to about 850 L, about 800 L to about 900 L, about 800 L to about 950 L, about 800 L to about 1,000 L, about 800 L to about 2,000 L, about 800 L to about 3,000 L, about 800 L to about 4,000 L, about 800 L to about 5,000 L, about 800 L to about 6,000 L, about 800 L to about 7,000 L, about 800 L to about 8,000 L, about 800 L to about 9,000 L, about 800 L to 10,000 L; about 900 L to about 950 L, about 900 L to about 1,000 L, about 900 L to about 2,000 L, about 900 L to about 3,000 L, about 900 L to about 4,000 L, about 900 L to about 5,000 L, about 900 L to about 6,000 L, about 900 L to about 7,000 L, about 900 L to about 8,000 L, about 900 L to about 9,000 L, about 900 L to 10,000 L; about 950 L to about 10,000 L). Vectors

Provided herein are cells in a liquid cell culture that can include a nucleic acid or a vector encoding a recombinant protein (e.g., any of the recombinant proteins described herein). The nucleic acid that encodes any of the recombinant proteins described herein can be included in an expression vector. Skilled practitioners will be capable of selecting suitable expression vectors for making the recombinant proteins described herein, and using the expression vectors to express a recombinant protein in a cell. For example, in selecting an expression vector, the cell must be considered because the vector may need to be able to replicate in it and/or be able to integrate into a chromosome of the cell. Exemplary expression vectors that can be used to produce a recombinant protein are also described below.

A“vector” according to the present disclosure is a polynucleotide capable of inducing the expression of a recombinant protein (e.g., any of the recombinant proteins as described herein) in a cell. A vector functions like a“molecular carrier,” delivering fragments of nucleic acids into a host cell (e.g., any of the cell described herein). It may include an expression cassette including regulatory sequences.

Foreign polynucleotides may be inserted into the expression cassette(s) of the vector nucleic acid in order to be expressed therefrom. The vectors according to the present invention may be present in circular or linearized form. The term“vector” also includes artificial chromosomes or similar respective polynucleotides allowing the transfer of foreign nucleic acid fragments. A respective vector can be used as expression vector in order to produce a recombinant protein as described herein in a cell.

Methods for constructing appropriate vectors, carrying DNA that codes for the recombinant protein described herein, and suitable for transforming (e.g., eukaryotic cells, mammalian cells, insect cells, yeast cells, bacterial cells) are well known in the art. See, for example, Ausubel et al., Eds.“Current Protocols in Molecular Biology” Current Protocols, 1993; and Sambrook et al., Eds.“Molecular Cloning: A Laboratory Manual,” 2nd ed., Cold Spring Harbor Press, 1989. Methods of Introducing a Nucleic Acid into a Cell

A variety of different methods known in the art can be used to introduce any of the nucleic acids and expression vectors disclosed herein into a cell (e.g., a eukaryotic cell, a plant cell). Non-limiting examples of methods that can be used to introduce a nucleic acid into a cell include lipofection, transfection, electroporation, microinjection, calcium phosphate transfection, dendrimer-based transfection, cationic polymer transfection, cell squeezing, sonoporation, transformation, optical transfection, impalection, hydrodynamic delivery, magnetofection, viral transduction (e.g., adenoviral and lentiviral transduction), and nanoparticle transfection. These and other methods of introducing a nucleic acid into a cell are well known in the art. In some embodiments, the introduced nucleic acid into a cell is transiently expressed. In some embodiments, the introduced nucleic acid into a cell is integrated into the cell genome. Contacting a Pod with a Liquid Cell Culture

The contacting step results in at least a portion (e.g., at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%) of the at least one area contacting the liquid cell culture. In some embodiments, the contacting is performed under conditions that result in the capture of the recombinant protein by the chromatography resin. In some embodiments, the contacting is performed under sterile conditions. In some embodiments, the contacting can be performed during agitation of the liquid cell culture (e.g., by rocking, by tilting, by rotary agitation (e.g., using a propeller), by vortexing, and/or aeration). In some embodiments, the contacting can occur when a liquid cell culture is in its exponential growth phase or stationary growth phase. In some embodiments, the contacting can occur when a liquid cell culture is in its death phase. In some embodiments of any of the methods described herein, the contacting can occur in a clarified supernatant of a liquid cell culture.

In some embodiments, the contacting is performed at a temperature of about 20 °C to about 40 °C (e.g., about 20 °C to about 40 °C, about 20 °C to about 25 °C, about 20 °C to about 30 °C, about 20 °C to about 35 °C, about 20 °C to about 37 °C, about 20 °C to about 38 °C, about 20 °C to about 39 °C, about 27 °C to about 30 °C, about 25 °C to about 30 °C, about 25 °C to about 37 °C, about 25 °C to about 38 °C, about 25 °C to about 39 °C, about 25 °C to about 40 °C, about 27 °C to about 30 °C, about 27 °C to about 35 °C, about 27 °C to about 37 °C, about 27 °C to about 38 °C, about 27 °C to about 39 °C, about 30 °C to about 35 °C, about 30 °C to about 37 °C, about 30 °C to about 38 °C, about 30 °C to about 40 °C, about 35 °C to about 38 °C, or about 32 °C to about 38 °C).

In some embodiments, the contacting is performed at a temperature of about 4 °C to about 18 °C (e.g., about 4 °C to about 16 °C, about 4 °C to about 14 °C, about 4 °C to about 12 °C about 4 °C to about 10 °C about 4 °C to about 8 °C about 4 °C to about 6 °C, about 6 °C to about 18 °C, about 6 °C to about 16 °C, about 6 °C to about 14 °C, about 6 °C to about 12 °C about 6 °C to about 10 °C, about 6 °C to about 8 °C, about 8 °C to about 18 °C, about 8 °C to about 16 °C, about 8 °C to about 14 °C, about 8 °C to about 12 °C, about 8 °C to about 10 °C, about 10 °C to about 18 °C, about 10 °C to about 16 °C, about 10 °C to about 14 °C, about 10 °C to about 12 °C, about 12 °C to about 18 °C, about 12 °C to about 16 °C, about 12 °C to about 14 °C, about 14 °C to about 18 °C, about 14 °C to about 16 °C, or about 16 °C to about 18 °C,).

In some embodiments, contacting can occur when the pod is at a 90° angle ± 50 ^o (e.g., at a 50 ^o to 140° angle, at a 50 ^o to 135° angle, at a 50 ^o to 130° angle, at a 50 ^o to 125° angle, at a 50 ^o to 120° angle, at a 50 ^o to 115° angle, at a 50 ^o to 110° angle, at a 50 ^o to 105° angle, at a 50 ^o to 100° angle, at a 50 ^o to 95° angle, at a 50 ^o to 90° angle, at a 50 ^o to 85° angle, at a 50 ^o to 80° angle, at a 50 ^o to 75° angle, at a 50 ^o to 70° angle, at a 50 ^o to 65° angle, at a 50 ^o to 60° angle, at a 55 ^o to 140° angle, at a 55 ^o to 135° angle, at a 55 ^o to 130° angle, at a 55 ^o to 125° angle, at a 55 ^o to 120° angle, at a 55 ^o to 115 ^o angle, at a 55 ^o to 110° angle, at a 55 ^o to 105° angle, at a 55 ^o to 100° angle, at a 55 ^o to 95° angle, at a 55 ^o to 90° angle, at a 55 ^o to 85° angle, at a 55 ^o to 80° angle, at a 55 ^o to 75° angle, at a 55 ^o to 70° angle, at a 55 ^o to 65° angle, at a 55 ^o to 60° angle, at a 60 ^o to 140° angle, at a 60 ^o to 135° angle, at a 60 ^o to 130° angle, at a 60 ^o to 125° angle, at a 60 ^o to 120° angle, at a 60 ^o to 115° angle, at a 60 ^o to 110° angle, at a 60 ^o to 105° angle, at a 60 ^o to 100° angle, at a 60 ^o to 95° angle, at a 60 ^o to 90° angle, at a 60 ^o to 85° angle, at a 60 ^o to 80° angle, at a 60 ^o to 75° angle, at a 60 ^o to 70° angle, at a 60 ^o to 65° angle, at a 65 ^o to 140° angle, at a 65 ^o to 135° angle, at a 65 ^o to 130° angle, at a 65 ^o to 125 ^o angle, at a 65 ^o to 120° angle, at a 65 ^o to 115° angle, at a 65 ^o to 110° angle, at a 65 ^o to 105° angle, at a 65 ^o to 100° angle, at a 65 ^o to 95° angle, at a 65 ^o to 90° angle, at a 65 ^o to 85° angle, at a 65 ^o to 80° angle, at a 65 ^o to 75° angle, at a 65 ^o to 70° angle, at a 70 ^o to 140° angle, at a 70 ^o to 135° angle, at a 70 ^o to 130° angle, at a 70 ^o to 125° angle, at a 70 ^o to 120° angle, at a 70 ^o to 115° angle, at a 70 ^o to 110° angle, at a 70 ^o to 105° angle, at a 70 ^o to 100 ^o angle, at a 70 ^o to 95 ^o angle, at a 70 ^o to 90 ^o angle, at a 70 ^o to 85 ^o angle, at a 70 ^o to 80 ^o angle, at a 70 ^o to 75 ^o angle, at a 75 ^o to 140 ^o angle, at a 75 ^o to 135 ^o angle, at a 75 ^o to 130 ^o angle, at a 75 ^o to 125 ^o angle, at a 75 ^o to 120 ^o angle, at a 75 ^o to 115 ^o angle, at a 75 ^o to 110 ^o angle, at a 75 ^o to 105 ^o angle, at a 75 ^o to 100 ^o angle, at a 75 ^o to 95 ^o angle, at a 75 ^o to 90 ^o angle, at a 75 ^o to 85 ^o angle, at a 75 ^o to 80 ^o angle, at a 80 ^o to 140 ^o angle, at a 80 ^o to 135 ^o angle, at a 80 ^o to 130 ^o angle, at a 80 ^o to 125 ^o angle, at a 80 ^o to 120 ^o angle, at a 80 ^o to 115 ^o angle, at a 80 ^o to 110 ^o angle, at a 80 ^o to 105 ^o angle, at a 80 ^o to 100 ^o angle, at a 80 ^o to 95 ^o angle, at a 80 ^o to 90 ^o angle, at a 80 ^o to 85 ^o angle, at a 85 ^o to 140 ^o angle, at a 85 ^o to 135 ^o angle, at a 85 ^o to 130 ^o angle, at a 85 ^o to 125 ^o angle, at a 85 ^o to 120 ^o angle, at a 85 ^o to 115 ^o angle, at a 85 ^o to 110 ^o angle, at a 85 ^o to 105 ^o angle, at a 85 ^o to 100 ^o angle, at a 85 ^o to 95 ^o angle, at a 85 ^o to 90 ^o angle, at a 90 ^o to 140 ^o angle, at a 90 ^o to 135 ^o angle, at a 90 ^o to 130 ^o angle, at a 90 ^o to 125 ^o angle, at a 90 ^o to 120 ^o angle, at a 90 ^o to 115 ^o angle, at a 90 ^o to 110 ^o angle, at a 90 ^o to 105 ^o angle, at a 90 ^o to 100 ^o angle, at a 90 ^o to 95 ^o angle, at a 95 ^o to 140 ^o angle, at a 95 ^o to 135 ^o angle, at a 95 ^o to 130 ^o angle, at a 95 ^o to 125 ^o angle, at a 95 ^o to 120 ^o angle, at a 95 ^o to 115 ^o angle, at a 95 ^o to 110 ^o angle, at a 95 ^o to 105 ^o angle, at a 95 ^o to 100 ^o angle, at a 100 ^o to 140 ^o angle, at a 100 ^o to 135 ^o angle, at a 100 ^o to 130 ^o angle, at a 100 ^o to 125 ^o angle, at a 100 ^o to 120 ^o angle, at a 100 ^o to 115 ^o angle, at a 100 ^o to 110 ^o angle, at a 100 ^o to 105 ^o angle, at a 105 ^o to 140 ^o angle, at a 105 ^o to 135 ^o angle, at a 105 ^o to 130 ^o angle, at a 105 ^o to 125 ^o angle, at a 105 ^o to 120 ^o angle, at a 105 ^o to 115 ^o angle, at a 105 ^o to 110 ^o angle, at a 110 ^o to 140 ^o angle, at a 110 ^o to 135 ^o angle, at a 110 ^o to 130 ^o angle, at a 110 ^o to 125 ^o angle, at a 110 ^o to 120 ^o angle, at a 110 ^o to 115 ^o angle, at a 115 ^o to 140 ^o angle, at a 115 ^o to 135 ^o angle, at a 115 ^o to 130 ^o angle, at a 115 ^o to 125 ^o angle, at a 115 ^o to 120 ^o angle, at a 120 ^o to 140 ^o angle, at a 120 ^o to 135 ^o angle, at a 120 ^o to 130 ^o angle, at a 120 ^o to 125 ^o angle, at a 125 ^o to 140 ^o angle, at a 125 ^o to 135 ^o angle, at a 125 ^o to 130 ^o angle, at a 130 ^o to 140 ^o angle, at a 130 ^o to 135 ^o angle, at a 135 ^o to 140 ^o angle, at a 50 ^o angle, at a 55 ^o angle, at a 60 ^o angle, at a 65 ^o angle, at a 70 ^o angle, at a 75 ^o angle, at a 80 ^o angle, at a 85 ^o angle, at a 90 ^o angle, at a 95 ^o angle, at a 100 ^o angle, at a 105 ^o angle, at a 110 ^o angle, at a 115 ^o angle, at a 120 ^o angle, at a 125 ^o angle, at a 130 ^o angle, at a 135 ^o angle, at a 140 ^o angle) from horizontal (e.g., from the bottom of a culture vessel (e.g., any of the culture vessels described herein)).

In some embodiments, contacting can occur when the pod is at a 180 ^o angle ± 35 ^o (e.g., at a 145 ^o to 180 ^o angle, at a 145 ^o to 175 ^o angle, at a 145 ^o to 170 ^o angle, at a 145 ^o to 165 ^o angle, at a 145 ^o to 160 ^o angle, at a 145 ^o to 155 ^o angle, at a 145 ^o to 150 ^o angle, at a 150 ^o to 180 ^o angle, at a 150 ^o to 175 ^o angle, at a 150 ^o to 170 ^o angle, at a 150 ^o to 165 ^o angle, at a 150 ^o to 160 ^o angle, at a 150 ^o to 155 ^o angle, at a 155 ^o to 180 ^o angle, at a 155 ^o to 175 ^o angle, at a 155 ^o to 170 ^o angle, at a 155 ^o to 165 ^o angle, at a 155 ^o to 160 ^o angle, at a 160 ^o to 180 ^o angle, at a 160 ^o to 175 ^o angle, at a 160 ^o to 170 ^o angle, at a 160 ^o to 165 ^o angle, at a 165 ^o to 180 ^o angle, at a 165 ^o to 175 ^o angle, at a 165 ^o to 170 ^o angle, at a 170 ^o to 180 ^o angle, at a 170 ^o to 175 ^o angle, at a 175 ^o to 180 ^o angle, at an angle 145 ^o , at an angle 150 ^o , at an angle 155 ^o , at an angle 160 ^o , at an angle 165 ^o , at an angle 170 ^o , at an angle 175 ^o , at an angle 180 ^o ) from horizontal (e.g., from the bottom of a culture vessel (e.g., any of the culture vessels described herein)). Sterilizing a Liquid Culture Medium and/or a Pod including a Chromatography Resin

In some embodiments, a pod including a chromatography resin can be sterilized. Non-limiting examples of methods of sterilization include: filtration, gamma-irradiation, electron-beam radiation, steam sterilization, autoclaving, and chemical sterilization (e.g., hydrogen chloride, sodium-N-lauroyl sarcosinate, sodium hydroxide, or ethanol).

In some embodiments, the chromatography resin is sterile. The term“sterile assurance level” as used herein refers to the probability of a single unit (e.g., a chromatography resin) being non-sterile after it has been subjected to a sterilization process. In some embodiments, the chromatography resin has a sterility assurance level of about 1 x 10 ^-5 to about 1 x 10 ^-15 (e.g., about 1 x 10 ^-5 to about 1 x 10 ^-10 , about 1 x 10 ^-5 to about 1 x 10 ^-15 , about 1 x 10 ^-6 to about 1 x 10 ^-10 , about 1 x 10 ^-6 to about 1 x 10 ^-12 , about 1 x 10 ^-6 to about 1 x 10 ^-15 , about 1 x 10 ^-5 , about 1 x 10 ^-6 , about 1 x 10 ^-7 , about 1 x 10 ^-7 , about 1 x 10 ^-8 , about 1 x 10 ^-9 , about 1 x 10 ^-10 , about 1 x 10 ^-11 , about 1 x 10 ^-12 , about 1 x 10 ^-13 , about 1 x 10 ^-14 , about 1 x 10 ^-15 ).

In some embodiments, the liquid cell culture medium present in the liquid cell culture can be sterile and/or sterilized. Washing Chromatography Resin

In some embodiments of any of the methods described herein, the method can further include a washing step. In some embodiments, the washing step is performed in between the contacting step and the eluting step. In some embodiments, the washing step includes one or more washes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) using a wash buffer. In some embodiments, the washing step removes or substantially removes contaminants and/or non-specific proteins. In some embodiments, the washing step equilibrates the chromatography resin of the pod before it is contacted with a liquid cell culture.

In some embodiments, the washing step is performed before removing the pod from the liquid cell culture and/or the culture vessel. In some embodiments, the washing step is performed after removing the pod from the liquid cell culture and/or the culture vessel. In some embodiments, the washing step is performed after removing the pod from the liquid cell culture and/or the culture vessel, and can involve pipetting a wash buffer directly into the interior of the pod.

In some embodiments the washing step is performed at about 2 °C to about 25 °C (e.g., 2 °C to about 4 °C, about 2 °C to about 6 °C, about 2 °C to about 8 °C, about 2 °C to about 10 °C, about 2 °C to about 12 °C, about 4 °C to about 6 °C, about 4 °C to about 8 °C, about 4 °C to about 10 °C, about 4 °C to about 12 °C, about 6 °C to about 8 °C, about 6 °C to about 10 °C, about 6 °C to about 12 °C, about 10 °C to about 12 °C, about 10 °C to about 25 °C, about 10 °C to about 22 °C, about 10 °C to about 20 °C, about 10 °C to about 18 °C, about 10 °C to about 14 °C, about 12 °C to about 14 °C, about 12 °C to about 16 °C, about 16 °C to about 18 °C, about 16 °C to about 20 °C, about 16 °C to about 25 °C, about 20 °C to about 22 °C, about 20 °C to about 25 °C, or about 22 °C to about 25 °C).

In some embodiments, the washing step is performed using a wash buffer that contains about 2 mM to about 6 mM (e.g., about 2 mM to about 5 mM, about 2 mM to about 4 mM, about 2 mM to about 3 mM, about 3 mM to about 6 mM, about 3 mM to about 5 mM, about 3 mM to about 4 mM, about 4 mM to about 6 mM, about 4 mM to about 5 mM, about 5 mM to about 6 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM) of ammonium acetate. In some embodiments, the wash buffer can further include salts and/or detergents. As is known in the art, the washing buffer is not meant to dissociate the recombinant protein from the chromatography resin.

In some embodiments, the washing step is performed using about 0.05 mL to about 1,000 mL, about 0.05 mL to about 950 mL, about 0.05 mL to about 900 mL, about 0.05 mL to about 850 mL, about 0.05 mL to about 800 mL, about 0.05 mL to about 750 mL, about 0.05 mL to about 700 mL, about 0.05 mL to about 650 mL, about 0.05 mL to about 600 mL, about 0.05 mL to about 550 mL, about 0.05 mL to about 500 mL, about 0.05 mL to about 450 mL, about 0.05 mL to about 400 mL, about 0.05 mL to about 350 mL, about 0.05 mL to about 300 mL, about 0.05 mL to about 250 mL, about 0.05 mL to about 200 mL, about 0.05 mL to about 150 mL, about 0.05 mL to about 100 mL, about 0.05 mL to about 90 mL, about 0.05 mL to about 80 mL, about 0.05 mL to about 70 mL, about 0.05 mL to about 60 mL, about 0.05 mL to about 50 mL, about 0.05 mL to about 45 mL, about 0.05 mL to about 40 mL, about 0.05 mL to about 35 mL, about 0.05 mL to about 30 mL, about 0.05 mL to about 25 mL, about 0.05 mL to about 20 mL, about 0.05 mL to about 15 mL, about 0.05 mL to about 10 mL, about 0.05 mL to about 8 mL, about 0.05 mL to about 6 mL, about 0.05 mL to about 5 mL, about 0.05 mL to about 4 mL, about 0.05 mL to about 3 mL, about 0.05 mL to about 2 mL, about 0.05 mL to about 1 mL, about 0.05 mL to about 0.5 mL, about 0.05 mL to about 0.1 mL, about 0.1 mL to about 1,000 mL, about 0.1 mL to about 950 mL, about 0.1 mL to about 900 mL, about 0.1 mL to about 850 mL, about 0.1 mL to about 800 mL, about 0.1 mL to about 750 mL, about 0.1 mL to about 700 mL, about 0.1 mL to about 650 mL, about 0.1 mL to about 600 mL, about 0.1 mL to about 550 mL, about 0.1 mL to about 500 mL, about 0.1 mL to about 450 mL, about 0.1 mL to about 400 mL, about 0.1 mL to about 350 mL, about 0.1 mL to about 300 mL, about 0.1 mL to about 250 mL, about 0.1 mL to about 200 mL, about 0.1 mL to about 150 mL, about 0.1 mL to about 100 mL, about 0.1 mL to about 90 mL, about 0.1 mL to about 80 mL, about 0.1 mL to about 70 mL, about 0.1 mL to about 60 mL, about 0.1 mL to about 50 mL, about 0.1 mL to about 45 mL, about 0.1 mL to about 40 mL, about 0.1 mL to about 35 mL, about 0.1 mL to about 30 mL, about 0.1 mL to about 25 mL, about 0.1 mL to about 20 mL, about 0.1 mL to about 15 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 8 mL, about 0.1 mL to about 6 mL, about 0.1 mL to about 5 mL, about 0.1 mL to about 4 mL, about 0.1 mL to about 3 mL, about 0.1 mL to about 2 mL, about 0.1 mL to about 1 mL, about 0.1 mL to about 0.5 mL, about 0.5 mL to about 1,000 mL, about 0.5 mL to about 950 mL, about 0.5 mL to about 900 mL, about 0.5 mL to about 850 mL, about 0.5 mL to about 800 mL, about 0.5 mL to about 750 mL, about 0.5 mL to about 700 mL, about 0.5 mL to about 650 mL, about 0.5 mL to about 600 mL, about 0.5 mL to about 550 mL, about 0.5 mL to about 500 mL, about 0.5 mL to about 450 mL, about 0.5 mL to about 400 mL, about 0.5 mL to about 350 mL, about 0.5 mL to about 300 mL, about 0.5 mL to about 250 mL, about 0.5 mL to about 200 mL, about 0.5 mL to about 150 mL, about 0.5 mL to about 100 mL, about 0.5 mL to about 90 mL, about 0.5 mL to about 80 mL, about 0.5 mL to about 70 mL, about 0.5 mL to about 60 mL, about 0.5 mL to about 50 mL, about 0.5 mL to about 45 mL, about 0.5 mL to about 40 mL, about 0.5 mL to about 35 mL, about 0.5 mL to about 30 mL, about 0.5 mL to about 25 mL, about 0.5 mL to about 20 mL, about 0.5 mL to about 15 mL, about 0.5 mL to about 10 mL, about 0.5 mL to about 8 mL, about 0.5 mL to about 6 mL, about 0.5 mL to about 5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1 mL, 1 mL to about 1 L, about 1 mL to about 900 mL, about 1 mL to about 800 mL, about 1 mL to about 700 mL, about 1 mL to about 600 mL, about 1 mL to about 500 mL, about 1 mL to about 250 mL, about 1 mL to about 400 mL, about 1 mL to about 350 mL, about 1 mL to about 300 mL, about 1 mL to about 250 mL, about 1 mL to about 200 mL, about 1 mL to about 150 mL, about 1 mL to about 100 mL, about 1 mL to about 90 mL, about 1 mL to about 80 mL, about 1 mL to about 70 mL, about 1 mL to about 60 mL, about 1 mL to about 50 mL, about 1 mL to about 40 mL, about 1 mL to about 30 mL, about 1 mL to about 20 mL, about 1 mL to about 10 mL, about 1 mL to about 5 mL; about 2 mL to about 1 L, about 2 mL to about 900 mL, about 2 mL to about 800 mL, about 2 mL to about 700 mL, about 2 mL to about 600 mL, about 2 mL to about 500 mL, about 2 mL to about 250 mL, about 2 mL to about 400 mL, about 2 mL to about 350 mL, about 2 mL to about 300 mL, about 2 mL to about 250 mL, about 2 mL to about 200 mL, about 2 mL to about 150 mL, about 2 mL to about 100 mL, about 2 mL to about 90 mL, about 2 mL to about 80 mL, about 2 mL to about 70 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 40 mL, about 2 mL to about 30 mL, about 2 mL to about 20 mL, about 2 mL to about 10 mL, about 2 mL to about 5 mL; about 5 mL to about 1 L, about 5 mL to about 900 mL, about 5 mL to about 800 mL, about 5 mL to about 700 mL, about 5 mL to about 600 mL, about 5 mL to about 500 mL, about 5 mL to about 250 mL, about 5 mL to about 400 mL, about 5 mL to about 350 mL, about 5 mL to about 300 mL, about 5 mL to about 250 mL, about 5 mL to about 200 mL, about 5 mL to about 150 mL, about 5 mL to about 100 mL, about 5 mL to about 90 mL, about 5 mL to about 80 mL, about 5 mL to about 70 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 40 mL, about 5 mL to about 30 mL, about 5 mL to about 20 mL, about 5 mL to about 10 mL; about 10 mL to about 1 L, about 10 mL to about 900 mL, about 10 mL to about 800 mL, about 10 mL to about 700 mL, about 10 mL to about 600 mL, about 10 mL to about 500 mL, about 10 mL to about 450 mL, about 10 mL to about 400 mL, about 10 mL to about 350 mL, about 10 mL to about 300 mL, about 10 mL to about 250 mL, about 10 mL to about 200 mL, about 10 mL to about 150 mL, about 10 mL to about 100 mL, about 10 mL to about 90 mL, about 10 mL to about 80 mL, about 10 mL to about 70 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 40 mL, about 10 mL to about 30 mL, about 10 mL to about 20 mL; about 20 mL to about 1 L, about 20 mL to about 900 mL, about 20 mL to about 800 mL, about 20 mL to about 700 mL, about 20 mL to about 600 mL, about 20 mL to about 500 mL, about 20 mL to about 450 mL, about 20 mL to about 400 mL, about 20 mL to about 350 mL, about 20 mL to about 300 mL, about 20 mL to about 250 mL, about 20 mL to about 200 mL, about 20 mL to about 150 mL, about 20 mL to about 100 mL, about 20 mL to about 90 mL, about 20 mL to about 80 mL, about 20 mL to about 70 mL, about 20 mL to about 60 mL, about 20 mL to about 50 mL, about 20 mL to about 40 mL, about 20 mL to about 30 mL; about 50 mL to about 1 L, about 50 mL to about 900 mL, about 50 mL to about 800 mL, about 50 mL to about 700 mL, about 50 mL to about 600 mL, about 50 mL to about 500 mL, about 50 mL to about 450 mL, about 50 mL to about 400 mL, about 50 mL to about 350 mL, about 50 mL to about 300 mL, about 50 mL to about 250 mL, about 50 mL to about 200 mL, about 50 mL to about 150 mL, about 50 mL to about 100 mL, about 50 mL to about 90 mL, about 50 mL to about 80 mL, about 50 mL to about 70 mL, about 50 mL to about 60 mL; about 100 mL to about 1 L, about 100 mL to about 900 mL, about 100 mL to about 800 mL, about 100 mL to about 700 mL, about 100 mL to about 600 mL, about 100 mL to about 500 mL, about 100 mL to about 450 mL, about 100 mL to about 400 mL, about 100 mL to about 350 mL, about 100 mL to about 300 mL, about 100 mL to about 250 mL, about 100 mL to about 200 mL, about 100 mL to about 150 mL; about 150 mL to about 1 L, about 150 mL to about 900 mL, about 150 mL to about 800 mL, about 150 mL to about 700 mL, about 150 mL to about 600 mL, about 150 mL to about 500 mL, about 150 mL to about 250 mL, about 150 mL to about 400 mL, about 150 mL to about 350 mL, about 150 mL to about 300 mL, about 150 mL to about 250 mL, about 150 mL to about 200 mL; about 200 mL to about 1 L, about 200 mL to about 900 mL, about 200 mL to about 800 mL, about 200 mL to about 700 mL, about 200 mL to about 600 mL, about 200 mL to about 500 mL, about 200 mL to about 450 mL, about 200 mL to about 400 mL, about 200 mL to about 350 mL, about 200 mL to about 300 mL, about 200 mL to about 250 mL; about 250 mL to about 1 L, about 250 mL to about 900 mL, about 250 mL to about 800 mL, about 250 mL to about 700 mL, about 250 mL to about 600 mL, about 250 mL to about 500 mL, about 250 mL to about 450 mL, about 250 mL to about 400 mL, about 250 mL to about 350 mL, about 250 mL to about 300 mL; about 300 mL to about 1 L, about 300 mL to about 900 mL, about 300 mL to about 800 mL, about 300 mL to about 700 mL, about 300 mL to about 600 mL, about 300 mL to about 500 mL, about 300 mL to about 450 mL, about 300 mL to about 400 mL, about 300 mL to about 350 mL; about 350 mL to about 1 L, about 350 mL to about 900 mL, about 350 mL to about 800 mL, about 350 mL to about 700 mL, about 350 mL to about 600 mL, about 350 mL to about 500 mL, about 350 mL to about 450 mL; about 350 mL to about 400 mL; about 400 mL to about 1 L, about 400 mL to about 900 mL, about 400 mL to about 800 mL, about 400 mL to about 700 mL, about 400 mL to about 600 mL, about 400 mL to about 500 mL, about 400 mL to about 450 mL; about 500 mL to about 1 L, about 500 mL to about 900 mL, about 500 mL to about 800 mL, about 500 mL to about 700 mL, about 500 mL to about 600 mL; about 550 mL to about 1 L, about 550 mL to about 900 mL, about 550 mL to about 800 mL, about 550 mL to about 700 mL, about 550 mL to about 600 mL; about 600 mL to about 1 L, about 600 mL to about 900 mL, about 600 mL to about 800 mL, about 600 mL to about 700 mL; about 600 mL to about 650 mL; about 650 mL to about 1 L, about 650 mL to about 900 mL, about 650 mL to about 800 mL, about 650 mL to about 700 mL; about 700 mL to about 1 L, about 700 mL to about 900 mL, about 700 mL to about 800 mL, about 700 mL to about 750 mL; about 750 mL to about 1 L, about 750 mL to about 900 mL, about 750 mL to about 800 mL; about 800 mL to about 1 L, about 800 mL to about 900 mL, about 800 mL to about 950 mL; about 850 mL to about 1 L, about 850 L to about 950 mL, about 850 mL to about 900 mL, about 900 mL to about 1 L, about 900 mL to about 950 mL) of a wash buffer (e.g., any of the wash buffers described herein).

In some embodiments of any of the methods described herein, the washing step can include one or more wash buffers (e.g., 2, 3, 4, 5, 6, 7, or 8) of any of the wash buffers described herein). Eluting a Recombinant Protein

In some embodiments, any of the pods described herein can be removed from the liquid culture medium before performing the eluting step. In some embodiments, the affinity chromatography resin of the pod is a protein A resin. In some embodiments where the affinity chromatography resin of the pod is a protein A resin, the eluting is performed using an elution buffer with an acidic pH. In some embodiments, the elution buffer includes arginine or citrate. In some embodiments, the elution buffer includes about 0.01 M to about 1 M (e.g., about 0.01 M to about 1 M, about 0.01 M to about 0.8 M, about 0.01 M to about 0.6 M, about 0.01 M to about 0.4 M, about 0.01 M to about 0.2 M, about 0.01 M to about 0.1 M, about 0.01 M to about 0.08 M, about 0.01 M to about 0.06 M, about 0.01 M to about 0.05 M, about 0.01 M to about 0.04 M, about 0.01 M to about 0.02 M; 0.02 M to about 1 M, about 0.02 M to about 0.8 M, about 0.02 M to about 0.6 M, about 0.02 M to about 0.4 M, about 0.02 M to about 0.2 M, about 0.02 M to about 0.1 M, about 0.02 M to about 0.08 M, about 0.02 M to about 0.06 M, about 0.02 M to about 0.05 M, about 0.02 M to about 0.04 M, 0.04 M to about 1 M, about 0.04 M to about 0.8 M, about 0.04 M to about 0.6 M, about 0.04 M to about 0.4 M, about 0.04 M to about 0.2 M, about 0.04 M to about 0.1 M, about 0.04 M to about 0.08 M, about 0.04 M to about 0.06 M, about 0.04 M to about 0.05 M, 0.05 M to about 1 M, about 0.05 M to about 0.8 M, about 0.05 M to about 0.6 M, about 0.05 M to about 0.4 M, about 0.05 M to about 0.2 M, about 0.05 M to about 0.1 M, about 0.05 M to about 0.08 M, about 0.05 M to about 0.06 M, 0.06 M to about 1 M, about 0.06 M to about 0.8 M, about 0.06 M to about 0.6 M, about 0.06 M to about 0.4 M, about 0.06 M to about 0.2 M, about 0.06 M to about 0.1 M, about 0.06 M to about 0.08 M, 0.08 M to about 1 M, about 0.08 M to about 0.8 M, about 0.08 M to about 0.6 M, about 0.08 M to about 0.4 M, about 0.08 M to about 0.2 M, about 0.08 M to about 0.1 M, 0.1 M to about 1 M, about 0.1 M to about 0.8 M, about 0.1 M to about 0.6 M, about 0.1 M to about 0.4 M, about 0.1 M to about 0.2 M,

0.2 M to about 1 M, about 0.2 M to about 0.8 M, about 0.2 M to about 0.6 M, about 0.2 M to about 0.4 M, 0.4 M to about 1 M, about 0.4 M to about 0.8 M, about 0.4 M to about 0.6 M, 0.6 M to about 1 M, about 0.6 M to about 0.8 M, 0.8 M to about 1 M, about 0.01 M, about 0.02 M, about 0.05 M, about 0.05 M, about 0.06 M, about 0.08 M, about 0.1 M, about 0.2 M, about 0.4 M, about 0.5 M, about 0.6 M, about 0.8 M, or about 1 M) of arginine or citrate. In some embodiments, the elution buffer includes about 0.1 to about 1 M (e.g., about 0.1 to about 0.9 M, about 0.1 M to about 0.8 M, about 0.1 M to about 0.7 M, about 0.1 M to about 0.6 M, about 0.1 M to about 0.5 M, about 0.1 M to about 0.4 M, about 0.1 M to about 0.3 M, about 0.1 M to about 0.2 M, about 0.2 M to about 1 M, about 0.2 M to about 0.9 M, about 0.2 M to about 0.8 M, about 0.2 M to about 0.7 M, about 0.2 M to about 0.6 M, about 0.2 M to about 0.5 M, about 0.2 M to about 0.4 M, about 0.2 M to about 0.3 M, about 0.3 M to about 1 M, about 0.3 M to about 0.9 M, about 0.3 M to about 0.8 M, about 0.3 M to about 0.7 M, about 0.3 M to about 0.6 M, about 0.3 M to about 0.5, about 0.3 M to about 0.4 M, about 0.4 M to about 1 M, about 0.4 M to about 0.9 M, about 0.4 to about 0.8 M, about 0.4 M to about 0.7 M, about 0.4 M to about 0.6 M, about 0.4 M to about 0.5 M, about 0.5 M to about 1 M, about 0.5 M to about 0.9 M, about 0.5 M to about 0.8 M, about 0.5 M to about 0.7 M, about 0.5 M to about 0.6 M, about 0.6 M to about 1 M, about 0.6 M to about 0.9 M, about 0.6 M to about 0.8 M, about 0.6 M to about 0.7 M, about 0.7 M to about 1 M, about 0.7 M to about 0.9 M, about 0.7 M to about 0.8 M, about 0.8 M to about 1 M, about 0.8 M to about 0.9 M, about 0.9 M to about 1 M) of acetic acid.

In some embodiments, the elution buffer includes about 0.1 M of glycine (pH 3.0) or about 0.1 M of citric acid (pH 3.0). In some embodiments of any of the chromatography resins used herein, the eluted recombinant proteins are neutralized using a buffer that contains about 1 M Tris (pH 9.0). In some embodiments of any of the chromatography resins used herein, the eluted recombinant proteins are neutralized using a buffer that contains about 1 M HEPES (pH 7.0) (e.g., between about 0.01 M to about 1 M, about 0.01 M to about 0.8 M, about 0.01 M to about 0.6 M, about 0.01 M to about 0.4 M, about 0.01 M to about 0.2 M, about 0.01 to about 0.1 M, about 0.01 M to about 0.05 M; about 0.02 M to about 1 M, about 0.02 M to about 0.8 M, about 0.02 M to about 0.6 M, about 0.02 M to about 0.4 M, about 0.02 M to about 0.2 M, about 0.02 to about 0.1 M, about 0.05 M to about 1 M, about 0.05 M to about 0.8 M, about 0.05 M to about 0.6 M, about 0.05 M to about 0.4 M, about 0.05 M to about 0.2 M, about 0.05 to about 0.1 M, about 0.1 M to about 1 M, about 0.1 M to about 0.8 M, about 0.1 M to about 0.6 M, about 0.1M to about 0.4 M, about 0.1 M to about 0.2 M, about 0.2 M to about 1 M, about 0.2 M to about 0.8 M, about 0.2 M to about 0.6 M, about 0.2 M to about 0.4 M, about 0.4 M to about 1 M, about 0.4 M to about 0.8 M, about 0.4 M to about 0.6 M, about 0.6 M to about 1 M, about 0.6 M to about 0.8 M, or about 0.8 M to about 1 M).

In some embodiments, the eluting is performed using about 0.05 mL to about 1,000 mL, about 0.05 mL to about 950 mL, about 0.05 mL to about 900 mL, about 0.05 mL to about 850 mL, about 0.05 mL to about 800 mL, about 0.05 mL to about 750 mL, about 0.05 mL to about 700 mL, about 0.05 mL to about 650 mL, about 0.05 mL to about 600 mL, about 0.05 mL to about 550 mL, about 0.05 mL to about 500 mL, about 0.05 mL to about 450 mL, about 0.05 mL to about 400 mL, about 0.05 mL to about 350 mL, about 0.05 mL to about 300 mL, about 0.05 mL to about 250 mL, about 0.05 mL to about 200 mL, about 0.05 mL to about 150 mL, about 0.05 mL to about 100 mL, about 0.05 mL to about 90 mL, about 0.05 mL to about 80 mL, about 0.05 mL to about 70 mL, about 0.05 mL to about 60 mL, about 0.05 mL to about 50 mL, about 0.05 mL to about 45 mL, about 0.05 mL to about 40 mL, about 0.05 mL to about 35 mL, about 0.05 mL to about 30 mL, about 0.05 mL to about 25 mL, about 0.05 mL to about 20 mL, about 0.05 mL to about 15 mL, about 0.05 mL to about 10 mL, about 0.05 mL to about 8 mL, about 0.05 mL to about 6 mL, about 0.05 mL to about 5 mL, about 0.05 mL to about 4 mL, about 0.05 mL to about 3 mL, about 0.05 mL to about 2 mL, about 0.05 mL to about 1 mL, about 0.05 mL to about 0.5 mL, about 0.05 mL to about 0.1 mL, about 0.1 mL to about 1,000 mL, about 0.1 mL to about 950 mL, about 0.1 mL to about 900 mL, about 0.1 mL to about 850 mL, about 0.1 mL to about 800 mL, about 0.1 mL to about 750 mL, about 0.1 mL to about 700 mL, about 0.1 mL to about 650 mL, about 0.1 mL to about 600 mL, about 0.1 mL to about 550 mL, about 0.1 mL to about 500 mL, about 0.1 mL to about 450 mL, about 0.1 mL to about 400 mL, about 0.1 mL to about 350 mL, about 0.1 mL to about 300 mL, about 0.1 mL to about 250 mL, about 0.1 mL to about 200 mL, about 0.1 mL to about 150 mL, about 0.1 mL to about 100 mL, about 0.1 mL to about 90 mL, about 0.1 mL to about 80 mL, about 0.1 mL to about 70 mL, about 0.1 mL to about 60 mL, about 0.1 mL to about 50 mL, about 0.1 mL to about 45 mL, about 0.1 mL to about 40 mL, about 0.1 mL to about 35 mL, about 0.1 mL to about 30 mL, about 0.1 mL to about 25 mL, about 0.1 mL to about 20 mL, about 0.1 mL to about 15 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 8 mL, about 0.1 mL to about 6 mL, about 0.1 mL to about 5 mL, about 0.1 mL to about 4 mL, about 0.1 mL to about 3 mL, about 0.1 mL to about 2 mL, about 0.1 mL to about 1 mL, about 0.1 mL to about 0.5 mL, about 0.5 mL to about 1,000 mL, about 0.5 mL to about 950 mL, about 0.5 mL to about 900 mL, about 0.5 mL to about 850 mL, about 0.5 mL to about 800 mL, about 0.5 mL to about 750 mL, about 0.5 mL to about 700 mL, about 0.5 mL to about 650 mL, about 0.5 mL to about 600 mL, about 0.5 mL to about 550 mL, about 0.5 mL to about 500 mL, about 0.5 mL to about 450 mL, about 0.5 mL to about 400 mL, about 0.5 mL to about 350 mL, about 0.5 mL to about 300 mL, about 0.5 mL to about 250 mL, about 0.5 mL to about 200 mL, about 0.5 mL to about 150 mL, about 0.5 mL to about 100 mL, about 0.5 mL to about 90 mL, about 0.5 mL to about 80 mL, about 0.5 mL to about 70 mL, about 0.5 mL to about 60 mL, about 0.5 mL to about 50 mL, about 0.5 mL to about 45 mL, about 0.5 mL to about 40 mL, about 0.5 mL to about 35 mL, about 0.5 mL to about 30 mL, about 0.5 mL to about 25 mL, about 0.5 mL to about 20 mL, about 0.5 mL to about 15 mL, about 0.5 mL to about 10 mL, about 0.5 mL to about 8 mL, about 0.5 mL to about 6 mL, about 0.5 mL to about 5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1 mL, 1 mL to about 1 L, about 1 mL to about 900 mL, about 1 mL to about 800 mL, about 1 mL to about 700 mL, about 1 mL to about 600 mL, about 1 mL to about 500 mL, about 1 mL to about 250 mL, about 1 mL to about 400 mL, about 1 mL to about 350 mL, about 1 mL to about 300 mL, about 1 mL to about 250 mL, about 1 mL to about 200 mL, about 1 mL to about 150 mL, about 1 mL to about 100 mL, about 1 mL to about 90 mL, about 1 mL to about 80 mL, about 1 mL to about 70 mL, about 1 mL to about 60 mL, about 1 mL to about 50 mL, about 1 mL to about 40 mL, about 1 mL to about 30 mL, about 1 mL to about 20 mL, about 1 mL to about 10 mL, about 1 mL to about 5 mL; about 2 mL to about 1 L, about 2 mL to about 900 mL, about 2 mL to about 800 mL, about 2 mL to about 700 mL, about 2 mL to about 600 mL, about 2 mL to about 500 mL, about 2 mL to about 250 mL, about 2 mL to about 400 mL, about 2 mL to about 350 mL, about 2 mL to about 300 mL, about 2 mL to about 250 mL, about 2 mL to about 200 mL, about 2 mL to about 150 mL, about 2 mL to about 100 mL, about 2 mL to about 90 mL, about 2 mL to about 80 mL, about 2 mL to about 70 mL, about 2 mL to about 60 mL, about 2 mL to about 50 mL, about 2 mL to about 40 mL, about 2 mL to about 30 mL, about 2 mL to about 20 mL, about 2 mL to about 10 mL, about 2 mL to about 5 mL; about 5 mL to about 1 L, about 5 mL to about 900 mL, about 5 mL to about 800 mL, about 5 mL to about 700 mL, about 5 mL to about 600 mL, about 5 mL to about 500 mL, about 5 mL to about 250 mL, about 5 mL to about 400 mL, about 5 mL to about 350 mL, about 5 mL to about 300 mL, about 5 mL to about 250 mL, about 5 mL to about 200 mL, about 5 mL to about 150 mL, about 5 mL to about 100 mL, about 5 mL to about 90 mL, about 5 mL to about 80 mL, about 5 mL to about 70 mL, about 5 mL to about 60 mL, about 5 mL to about 50 mL, about 5 mL to about 40 mL, about 5 mL to about 30 mL, about 5 mL to about 20 mL, about 5 mL to about 10 mL; about 10 mL to about 1 L, about 10 mL to about 900 mL, about 10 mL to about 800 mL, about 10 mL to about 700 mL, about 10 mL to about 600 mL, about 10 mL to about 500 mL, about 10 mL to about 450 mL, about 10 mL to about 400 mL, about 10 mL to about 350 mL, about 10 mL to about 300 mL, about 10 mL to about 250 mL, about 10 mL to about 200 mL, about 10 mL to about 150 mL, about 10 mL to about 100 mL, about 10 mL to about 90 mL, about 10 mL to about 80 mL, about 10 mL to about 70 mL, about 10 mL to about 60 mL, about 10 mL to about 50 mL, about 10 mL to about 40 mL, about 10 mL to about 30 mL, about 10 mL to about 20 mL; about 20 mL to about 1 L, about 20 mL to about 900 mL, about 20 mL to about 800 mL, about 20 mL to about 700 mL, about 20 mL to about 600 mL, about 20 mL to about 500 mL, about 20 mL to about 450 mL, about 20 mL to about 400 mL, about 20 mL to about 350 mL, about 20 mL to about 300 mL, about 20 mL to about 250 mL, about 20 mL to about 200 mL, about 20 mL to about 150 mL, about 20 mL to about 100 mL, about 20 mL to about 90 mL, about 20 mL to about 80 mL, about 20 mL to about 70 mL, about 20 mL to about 60 mL, about 20 mL to about 50 mL, about 20 mL to about 40 mL, about 20 mL to about 30 mL; about 50 mL to about 1 L, about 50 mL to about 900 mL, about 50 mL to about 800 mL, about 50 mL to about 700 mL, about 50 mL to about 600 mL, about 50 mL to about 500 mL, about 50 mL to about 450 mL, about 50 mL to about 400 mL, about 50 mL to about 350 mL, about 50 mL to about 300 mL, about 50 mL to about 250 mL, about 50 mL to about 200 mL, about 50 mL to about 150 mL, about 50 mL to about 100 mL, about 50 mL to about 90 mL, about 50 mL to about 80 mL, about 50 mL to about 70 mL, about 50 mL to about 60 mL; about 100 mL to about 1 L, about 100 mL to about 900 mL, about 100 mL to about 800 mL, about 100 mL to about 700 mL, about 100 mL to about 600 mL, about 100 mL to about 500 mL, about 100 mL to about 450 mL, about 100 mL to about 400 mL, about 100 mL to about 350 mL, about 100 mL to about 300 mL, about 100 mL to about 250 mL, about 100 mL to about 200 mL, about 100 mL to about 150 mL; about 150 mL to about 1 L, about 150 mL to about 900 mL, about 150 mL to about 800 mL, about 150 mL to about 700 mL, about 150 mL to about 600 mL, about 150 mL to about 500 mL, about 150 mL to about 250 mL, about 150 mL to about 400 mL, about 150 mL to about 350 mL, about 150 mL to about 300 mL, about 150 mL to about 250 mL, about 150 mL to about 200 mL; about 200 mL to about 1 L, about 200 mL to about 900 mL, about 200 mL to about 800 mL, about 200 mL to about 700 mL, about 200 mL to about 600 mL, about 200 mL to about 500 mL, about 200 mL to about 450 mL, about 200 mL to about 400 mL, about 200 mL to about 350 mL, about 200 mL to about 300 mL, about 200 mL to about 250 mL; about 250 mL to about 1 L, about 250 mL to about 900 mL, about 250 mL to about 800 mL, about 250 mL to about 700 mL, about 250 mL to about 600 mL, about 250 mL to about 500 mL, about 250 mL to about 450 mL, about 250 mL to about 400 mL, about 250 mL to about 350 mL, about 250 mL to about 300 mL; about 300 mL to about 1 L, about 300 mL to about 900 mL, about 300 mL to about 800 mL, about 300 mL to about 700 mL, about 300 mL to about 600 mL, about 300 mL to about 500 mL, about 300 mL to about 450 mL, about 300 mL to about 400 mL, about 300 mL to about 350 mL; about 350 mL to about 1 L, about 350 mL to about 900 mL, about 350 mL to about 800 mL, about 350 mL to about 700 mL, about 350 mL to about 600 mL, about 350 mL to about 500 mL, about 350 mL to about 450 mL; about 350 mL to about 400 mL; about 400 mL to about 1 L, about 400 mL to about 900 mL, about 400 mL to about 800 mL, about 400 mL to about 700 mL, about 400 mL to about 600 mL, about 400 mL to about 500 mL, about 400 mL to about 450 mL; about 500 mL to about 1 L, about 500 mL to about 900 mL, about 500 mL to about 800 mL, about 500 mL to about 700 mL, about 500 mL to about 600 mL; about 550 mL to about 1 L, about 550 mL to about 900 mL, about 550 mL to about 800 mL, about 550 mL to about 700 mL, about 550 mL to about 600 mL; about 600 mL to about 1 L, about 600 mL to about 900 mL, about 600 mL to about 800 mL, about 600 mL to about 700 mL; about 600 mL to about 650 mL; about 650 mL to about 1 L, about 650 mL to about 900 mL, about 650 mL to about 800 mL, about 650 mL to about 700 mL; about 700 mL to about 1 L, about 700 mL to about 900 mL, about 700 mL to about 800 mL, about 700 mL to about 750 mL; about 750 mL to about 1 L, about 750 mL to about 900 mL, about 750 mL to about 800 mL; about 800 mL to about 1 L, about 800 mL to about 900 mL, about 800 mL to about 950 mL; about 850 mL to about 1 L, about 850 L to about 950 mL, about 850 mL to about 900 mL, about 900 mL to about 1 L, or about 900 mL to about 950 mL) of an elution buffer (e.g., any of the elution buffers described herein).

In some embodiments, the eluting is performed at about 2 °C to about 25 °C (e.g., 2 °C to about 4 °C, about 2 °C to about 6 °C, about 2 °C to about 8 °C, about 2 °C to about 10 °C, about 2 °C to about 12 °C, about 4 °C to about 6 °C, about 4 °C to about 8 °C, about 4 °C to about 10 °C, about 4 °C to about 12 °C, about 6 °C to about 8 °C, about 6 °C to about 10 °C, about 6 °C to about 12 °C, about 10 °C to about 12 °C, about 10 °C to about 25 °C, about 10 °C to about 22 °C, about 10 °C to about 20 °C, about 10 °C to about 18 °C, about 10 °C to about 14 °C, about 12 °C to about 14 °C, about 12 °C to about 16 °C, about 16 °C to about 18 °C, about 16 °C to about 20 °C, about 16 °C to about 25 °C, about 20 °C to about 22 °C, about 20 °C to about 25 °C, about 22 °C to about 25 °C).

In some embodiments, the total amount of recombinant protein eluted is about 0.05 mg to about 80 mg (e.g., about 0.05 mg to about 0.1 mg, about 0.05 mg to about 0.2 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.6 mg, about 0.05 mg to about 0.8 mg, about 0.05 mg to about 1 mg, about 0.05 mg to about 2 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 15 mg, about 0.05 mg to about 20 mg, about 0.05 mg to about 25 mg, about 0.05 mg to about 30 mg, about 0.05 mg to about 35 mg, about 0.05 mg to about 40 mg, about 0.05 mg to about 45 mg, about 0.05 mg to about 50 mg, about 0.05 mg to about 55 mg, about 0.05 mg to about 60 mg, about 0.05 mg to about 65 mg, about 0.05 mg to about 70 mg, about 0.05 mg to about 75 mg, about 0.05 mg to about 80 mg, about 0.1 mg to about 0.2 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.6 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 1 mg, about 0.1 mg to about 2 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 10 mg, about 0.1 mg to about 15 mg, about 0.1 mg to about 20 mg, about 0.1 mg to about 25 mg, about 0.1 mg to about 30 mg, about 0.1 mg to about 35 mg, about 0.1 mg to about 40 mg, about 0.1 mg to about 45 mg, about 0.1 mg to about 50 mg, about 0.1 mg to about 55 mg, about 0.1 mg to about 60 mg, about 0.1 mg to about 65 mg, about 0.1 mg to about 70 mg, about 0.1 mg to about 75 mg, about 0.1 mg to about 80 mg, about 0.2 mg to about 0.5 mg, about 0.2 mg to about 0.6 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 1 mg, about 0.2 mg to about 2 mg, about 0.2 mg to about 5 mg, about 0.2 mg to about 10 mg, about 0.2 mg to about 15 mg, about 0.2 mg to about 20 mg, about 0.2 mg to about 25 mg, about 0.2 mg to about 30 mg, about 0.2 mg to about 35 mg, about 0.2 mg to about 40 mg, about 0.2 mg to about 45 mg, about 0.2 mg to about 50 mg, about 0.2 mg to about 55 mg, about 0.2 mg to about 60 mg, about 0.2 mg to about 65 mg, about 0.2 mg to about 70 mg, about 0.2 mg to about 75 mg, about 0.2 mg to about 80 mg, about 0.5 mg to about 0.6 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 1 mg, about 0.5 mg to about 2 mg, about 0.5 mg to about 5 mg, about 0.5 mg to about 10 mg, about 0.5 mg to about 15 mg, about 0.5 mg to about 20 mg, about 0.5 mg to about 25 mg, about 0.5 mg to about 30 mg, about 0.5 mg to about 35 mg, about 0.5 mg to about 40 mg, about 0.5 mg to about 45 mg, about 0.5 mg to about 50 mg, about 0.5 mg to about 55 mg, about 0.5 mg to about 60 mg, about 0.5 mg to about 65 mg, about 0.5 mg to about 70 mg, about 0.5 mg to about 75 mg, about 0.5 mg to about 80 mg, about 0.6 mg to about 0.8 mg, about 0.6 mg to about 1 mg, about 0.6 mg to about 2 mg, about 0.6 mg to about 5 mg, about 0.6 mg to about 10 mg, about 0.6 mg to about 15 mg, about 0.6 mg to about 20 mg, about 0.6 mg to about 25 mg, about 0.6 mg to about 30 mg, about 0.6 mg to about 35 mg, about 0.6 mg to about 40 mg, about 0.6 mg to about 45 mg, about 0.6 mg to about 50 mg, about 0.6 mg to about 55 mg, about 0.6 mg to about 60 mg, about 0.6 mg to about 65 mg, about 0.6 mg to about 70 mg, about 0.6 mg to about 75 mg, about 0.6 mg to about 80 mg, about 0.8 mg to about 1 mg, about 0.8 mg to about 2 mg, about 0.8 mg to about 5 mg, about 0.8 mg to about 10 mg, about 0.8 mg to about 15 mg, about 0.8 mg to about 20 mg, about 0.8 mg to about 25 mg, about 0.8 mg to about 30 mg, about 0.8 mg to about 35 mg, about 0.8 mg to about 40 mg, about 0.8 mg to about 45 mg, about 0.8 mg to about 50 mg, about 0.8 mg to about 55 mg, about 0.8 mg to about 60 mg, about 0.8 mg to about 65 mg, about 0.8 mg to about 70 mg, about 0.8 mg to about 75 mg, about 0.8 mg to about 80 mg, about 1 mg to about 2 mg, about 1 mg to about 5 mg, about 1 mg to about 10 mg, about 1 mg to about 15 mg, about 1 mg to about 20 mg, about 1 mg to about 25 mg, about 1 mg to about 30 mg, about 1 mg to about 35 mg, about 1 mg to about 40 mg, about 1 mg to about 45 mg, about 1 mg to about 50 mg, about 1 mg to about 55 mg, about 1 mg to about 60 mg, about 1 mg to about 65 mg, about 1 mg to about 70 mg, about 1 mg to about 75 mg, about 1 mg to about 80 mg, about 2 mg to about 5 mg, about 2 mg to about 10 mg, about 2 mg to about 15 mg, about 2 mg to about 20 mg, about 2 mg to about 25 mg, about 2 mg to about 30 mg, about 2 mg to about 35 mg, about 2 mg to about 40 mg, about 2 mg to about 45 mg, about 2 mg to about 50 mg, about 2 mg to about 55 mg, about 2 mg to about 60 mg, about 2 mg to about 65 mg, about 2 mg to about 70 mg, about 2 mg to about 75 mg, about 2 mg to about 80 mg, about 5 mg to about 10mg, about 5 mg to about 15 mg, about 5 mg to about 20 mg, about 5 mg to about 25 mg, about 5 mg to about 30 mg, about 5 mg to about 35 mg, about 5 mg to about 40 mg, about 5 mg to about 45 mg, about 5 mg to about 50 mg, about 5 mg to about 55 mg, about 5 mg to about 60 mg, about 5 mg to about 65 mg, about 5 mg to about 70 mg, about 5 mg to about 75 mg, about 5 mg to about 80 mg, about 10 mg to about 15 mg, about 10 mg to about 20 mg, about 10 mg to about 25 mg, about 10 mg to about 30 mg, about 10 mg to about 35 mg, about 10 mg to about 40 mg, about 10 mg to about 45 mg, about 10 mg to about 50 mg, about 10 mg to about 55 mg, about 10 mg to about 60 mg, about 10 mg to about 65 mg, about 10 mg to about 70 mg, about 10 mg to about 75 mg, about 10 mg to about 80 mg, about 15 mg to about 20 mg, about 15 mg to about 20 mg, about 15 mg to about 25 mg, about 15 mg to about 30 mg, about 15 mg to about 35 mg, about 15 mg to about 40 mg, about 15 mg to about 45 mg, about 15 mg to about 50 mg, about 15 mg to about 55 mg, about 15 mg to about 60 mg, about 15 mg to about 65 mg, about 15 mg to about 70 mg, about 15 mg to about 75 mg, about 15 mg to about 80 mg, about 20 mg to about 25 mg, about 20 mg to about 30 mg, about 20 mg to about 35 mg, about 20 mg to about 40 mg, about 20 mg to about 45 mg, about 20 mg to about 50 mg, about 20 mg to about 55 mg, about 20 mg to about 60 mg, about 20 mg to about 65 mg, about 20 mg to about 70 mg, about 20 mg to about 75 mg, about 20 mg to about 80 mg, about 25 mg to about 30 mg, about 25 mg to about 35 mg, about 25 mg to about 40 mg, about 25 mg to about 45 mg, about 25 mg to about 50 mg, about 25 mg to about 55 mg, about 25 mg to about 60 mg, about 25 mg to about 65 mg, about 25 mg to about 70 mg, about 25 mg to about 75 mg, about 25 mg to about 80 mg, about 30 mg to about 35 mg, about 30 mg to about 40 mg, about 30 mg to about 45 mg, about 30 mg to about 50 mg, about 30 mg to about 55 mg, about 30 mg to about 60 mg, about 30 mg to about 65 mg, about 30 mg to about 70 mg, about 30 mg to about 75 mg, about 30 mg to about 80 mg, about 35 mg to about 40 mg, about 35 mg to about 45 mg, about 35 mg to about 50 mg, about 35 mg to about 55 mg, about 35 mg to about 60 mg, about 35 mg to about 65 mg, about 35 mg to about 70 mg, about 35 mg to about 75 mg, about 35 mg to about 80 mg, about 40 mg to about 45 mg, about 40 mg to about 50 mg, about 45mg to about 55 mg, about 45 mg to about 60 mg, about 45 mg to about 65 mg, about 45 mg to about 70 mg, about 45 mg to about 75 mg, about 45 mg to about 80 mg, about 50 mg to about 55 mg, about 50 mg to about 60 mg, about 50 mg to about 65 mg, about 50 mg to about 70 mg, about 50 mg to about 75 mg, about 50 mg to about 80 mg, about 55 mg to about 60 mg, about 55 mg to about 65 mg, about 55 mg to about 70 mg, about 55 mg to about 75 mg, about 55 mg to about 80 mg, about 60 mg to about 65 mg, about 60 mg to about 70 mg, about 60 mg to about 75 mg, about 60 mg to about 80 mg, about 65 mg to about 70 mg, about 65 mg to about 75 mg, about 65 mg to about 80 mg, about 70 mg to about 75 mg, about 70 mg to about 80 mg, about 75 mg to about 80 mg). One or More Additional Unit Operations

In some embodiments of any of the methods provided herein, one or more additional unit operations can be performed on the recombinant protein. In some embodiments, the one or more additional unit operations is selected from the group of: viral inactivation, viral filtration, polishing, size-exclusion chromatography, cation exchange chromatography, anion exchange chromatography, hydrophobic interaction chromatography, mixed mode chromatography, reverse phase chromatography and ultrafiltration/diafiltration. In some embodiments, the one or more additional unit operations includes cleaning in place (CIP), where the precipitated and/or denatured substances are removed.

In some embodiments of any of the methods described herein, the method can further include a step of formulating the recombinant protein. In some embodiments, the recombinant protein is formulated into a recombinant protein drug product. EXAMPLES

The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. Example 1. Comparative Study of Traditional Purification Methods and “Capture & Purify” Method with Clarified Supernatant

Experimental Set-Up

Construct C26, a traditional IgG antibody, is expressed in HEK 293 suspension cells for 5 days at 1 L cell culture scale. Upon harvest of construct C26, the supernatant is clarified by centrifugation and by filtration through a filter with a pore size of 0.22 μm. The clarified supernatant is divided into four x 250 mL aliquots and stored at 4 ^o C until needed for testing. One hundred-fifty mL of the clarified supernatant (or 185 mL for ÄKTA purification) is used to test three different methods of purification. All three methods of purification are performed using the same elution buffer and the same neutralization buffer. The ÄKTA method uses a PBS wash and at least one citrate wash. The first purification method ÄKTA is a programmed purification method using an immobilized resin within a prepacked column (i.e. commercial column). The ÄKTA purification method is directional flow-based. The second purification method is a traditional batch capture method. This traditional batch capture method includes over-night shaking with the resin floating freely in the clarified supernatant. The third purification method, called “Capture & Purify”, includes the use of a chromatography filtration pod as described herein. The Capture & Purify method also includes over-night shaking identical to the traditional batch capture method. Results

Experiments revealed that the yield and purity of construct C26 was comparable across all three tested methods (Table 1). Table 1. Summary Table of Yield and Purity Comparing Three Methods of Purification

The ÄKTA purification method used a 5 mL resin, while both the traditional batch capture method and the Capture & Purify method used a 3 mL. Nevertheless, 3-mL of resin was sufficient to capture a comparable amount of IgG antibody in the supernatant based on the observed binding capacity of the resin. Next, SDS-PAGE was performed to analyze 20 μL of flow through (FT) samples, 20 μL of wash samples (W), 1 μg of elution sample (E) taken during the traditional batch purification method and the Capture & Purify method (Figure 5). Reduced and non-reduced samples of both purification methods were run on a Life Tech 4-12% gel. SDS analysis suggests that protein capture was complete in both methods, as the protein of interest was not lost in the flow through or during the wash steps of either method.

The experimental time line of the three methods is outlined in Figure 6. As shown in Figure 6, the ÄKTA purification method can take 7 days post-transfection to complete, with over 5-6 hours being dedicated to clarifying and setting up the experimental equipment. Furthermore, the ÄKTA purification method is restricted by flow rate, and thus for larger volumes (greater than 1 L), the time to completion can increase dramatically, and sometimes can take longer than indicated in Figure 6. In addition, only one protein can be purified at a time using the ÄKTA purification method. In contrast, the traditional batch purification method and the Capture & Purify method can capture and purify several proteins simultaneously in the timeframe as diagrammed in Figure 6. The traditional batch purification method can take 7 days post-transfection to complete, is the most labor-intensive method of the three tested methods. Both the ÄKTA purification method and the traditional batch method require far more consumables than the Capture and Purify method described herein. The Capture and Purify method is far less labor-intensive, and requires minimal user manipulation. The Capture and Purify method can be completed by day 6 post-transfection. Example 2. Study of“Capture & Purify” Method with a Pod including a Porous Mesh with an Average Pore Size of 20 µm or 40 µm

Experimental Set-Up

A 750 mL culture of a HIS tag human protein (C27), expressed as in example 1, was harvested and divided into two groups for post harvest processing. The supernatant marked“Filtered” in Table 2 was clarified by centrifugation and then passed through a filter with a 0.22 micron pore size. The other group was not processed in any way and is marked“unfiltered” in Table 2. Two one hundred twenty five mL aliquots of“Filtered” and“Unfiltered” culture media was added to 250 mL bottles. A Capture and Purify device with 1 ml of NI-NTA affinity resin and either 20 micron or 40 micron mesh covering the bottom of the device (the only opening in this instance) was added to each bottle. A traditional batch capture condition was not included in this experiment. Experimental conditions and yields are detailed in Table 2 below. Table 2. Experimental Conditions

As shown in Figures 7, high purity was seen across all four conditions (~92%). These data indicate that there was no loss in purity using a 40-μm porous mesh as compared to a 20-μm porous mesh and that the Capture and Purify device can obtain comparable yields to the traditional batch capture method using a HIS affinity tag purification. Example 3. Study of Purification Methods Using a Pod including a 40-μm Porous Mesh

Experimental Set-Up

Different experimental set-ups were tested for their ability to capture transiently expressed human IgG. In the first set-up (conditions #2 and 3), the pod included a porous mesh only at the bottom end of the pod. In the second set up (condition #4), the pod had a porous mesh on both the top and bottom ends of the pod with vents in the wall of the pod (FIG.4).

Both set ups were disposed in filtered (as described in Example 1 and 2) or unfiltered liquid cell culture, approximately 175 mL of culture media in a 150 mL bottle, by positioning the pod (in the filtered or unfiltered liquid cell culture) such that its longitudinal axis is approximately vertical. After the pod was removed from the liquid cell culture, the column was washed and then eluted using a PBS and citrate buffer, respectively. Table 3. Capture & Purify experimental conditions for IgG purification

The yields in mg/L are shown in Table 3 for all four conditions. While purity by HPLC was the same across all four conditions (Figure 8), the third and fourth set up had a higher yield as compared to the traditional batch capture culture, while the second set up was lower. Example 4. Study of Purification Methods Using a Pod having a 20-μm Porous Mesh

In this study, the traditional batch capture culture method using a filtered supernatant was compared to a method using an unfiltered supernatant and a pod as described in Example 3 that only had a porous mesh on the bottom end of the pod (Table 4). Table 4. Experimental conditions

The yields in mg/L are shown in Table 4 for both conditions. While purity by HPLC was the same across all both tested conditions (Figure 9), the Capture & Purify method had a higher yield as compared to the traditional batch capture culture. Example 5: Study of Purification Methods Using a Pod having 20 µm or 40 µm Porous Mesh and the addition of a stop cock to control the flow rate after absorption

In this study, a transient expression 0.9L of a IgG and subsequent purification was used to compare the traditional batch capture method to a method using a pod and unfiltered culture media. The pods, in this instance, had openings, which were covered by either 20 or 40 micron porous mesh, on their vertical sides and bases. The 0.9L culture was subdivided into 5 one hundred eighty mL aliquots. One of these aliquots was clarified by centrifugation and filtered through a filter with 0.22 micron pores, in the other four aliquots the cell culture media was not processed and termed “unfiltered”. All aliquots were added to 250 ml bottles. Pods, as described above and with 1.5 ml of protein A resin added to their interior were added to the unfiltered conditions and 1.5 ml of protein A resin was added to the filtered condition. All 250 ml bottles were then incubated overnight with shaking. Addition of a stopcock to control flow rate occurred after removal of the pod from the culture media following overnight incubation. See Table 5 for experimental conditions and results. Table 5: Experimental Conditions and Results

The yields are shown for all conditions. While purities are comparable for all conditions, the pods sometimes showed yields similar to the traditional batch capture method. Example 6. Study of Purification Methods Using a Pod including a 40-μm Porous Mesh

Experimental Set-Up

Different experimental set ups were tested for their ability to capture antibodies from either filtered or unfiltered but otherwise identical cell culture media containing an antibody. For set ups using a device as shown in Figs.27-28, the supernatant was unfiltered. For set ups, serving as a positive control, the supernatant was filtered prior to addition of the free floating resin.

The first set-up used 2 mL of free floating protein A chromatography resin exposed to filtered supernatant in free-floating batch capture environment with 150 RPM agitation. Subsequent purification employed a gravity flow column to capture the resin from the supernatant and then perform washes and elute the antibody from the resin. This gravity column was washed with 25 mL PBS, and eluted with 20 mL of elution buffer. This chromatography column was loaded with the same filtered cell culture medium containing an antibody as was used to contact the device/pod used in the third set-up.

The second set-up used 2.5 mL of free floating protein A chromatography resin exposed to filtered supernatant in a free-floating batch capture environment with 150 RPM agitation. Subsequent purification employed a gravity flow column to capture the resin from the supernatant and then perform washes and elute the antibody from the resin. This gravity column was washed with 37.5 mL of PBS and eluted with 30 mL of elution buffer. This chromatography column was loaded with the same filtered cell culture medium as was used to contact the device/pod used in the fourth set-up.

The third set-up used the device shown in Figures 27 and 28 loaded with 2-mL protein A chromatography resin. This resin in the pod/device was washed with 75 mL PBS, and eluted with 30 mL of elution buffer. After the elution buffer was first contacted with the resin in the pod/device, a 5 minute pause was implemented before additional elution buffer was flowed through the resin in the pod/device. The chromatography performed using an ÄKTA system. The resin in the pod/device was loaded with the same filtered cell culture medium as was used to contact the device/pod used in the first set-up. More specifically, the pod was contacted with 350 mL of the filtered cell culture medium in a 500 mL bottle.

The fourth set-up used the device shown in Figures 27 and 28 loaded with 3- mL protein A chromatography resin. This resin in the pod/device was washed with 65 mL PBS, and eluted with 25 mL of elution buffer (at a flow rate of 3 mL/min). The chromatography performed using an ÄKTA system. The resin in the pod/device was loaded with the same filtered cell culture medium as was used to contact the device/pod used in the second set-up. More specifically, the pod was contacted with 350 mL of the filtered cell culture medium in a 500 mL bottle. Results

The data are shown in Table 6 below. The chromatographs from the elution of the devices/pods from the third and the fourth set-ups are shown in Figures 32 and 33, respectively. Table 6. Capture & Purify experimental conditions for IgG purification

The above data show that the pods/devices provided herein provide similar yields of recombinant antibodies as compared to the use of standard chromatography columns. OTHER EMBODIMENTS

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.