PRIORITY

[0001] This application claims priority to U.S. Patent Application Serial No. 16/912,705, filed June 25, 2020.

BACKGROUND

[0002] The present disclosure relates to field flow fractionators, and more specifically, to sealing a field flow fractionator.

SUMMARY

[0003] The present disclosure describes a field flow fractionator. In an exemplary embodiment, the field flow fractionator includes a spacer including a core, and a coating covering the core. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate

(BoPET). In an embodiment, the plastic is polycarbonate. In an embodiment, the core includes stainless steel. In a particular embodiment, the core is stainless steel. In an embodiment, the coating is silicone. In an embodiment, the coating is polytetrafluoroethylene (PTFE). In an embodiment, the coating is ethylene tetrafluoroethylene (ETFE). In an alternative embodiment, the coating covers only the top side of the core. In an alternative embodiment, the coating covers only the bottom side of the core.

[0004] In an exemplary embodiment, the field flow fractionator includes (1) a spacer including a core, and a coating covering the core, (2) a bottom block including pins, and (3) atop block including mating holes to receive the pins, where the spacer is configured to be positioned between the top block and the bottom block, where the spacer includes mating holes to receive the pins. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET).

In an embodiment, the plastic is polycarbonate. In an embodiment, the coating is polytetrafluoroethylene (PTFE).

[0005] In an exemplary embodiment, the field flow fractionator includes (1) a spacer including (a) a core including an opening, atop side, and a bottom side, (b) a first coating covering the top side of the core, and (c) a second coating covering the bottom side of the core,

(2) a top block configured to press on the first coating along a first periphery of the opening, and

(3) a bottom block configured to press on the second coating along a second periphery of the opening. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET).

In an embodiment, the plastic is polycarbonate. In an embodiment, the first coating is polytetrafluoroethylene (PTFE). In an embodiment, the second coating is polytetrafluoroethylene (PTFE).

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] FIG. 1 depicts an existing field flow fractionator.

[0007] FIG. 2A depicts a field flow fractionator in accordance with an exemplary embodiment.

[0008] FIG. 2B depicts a field flow fractionator in accordance with an embodiment.

[0009] FIG. 2C depicts a field flow fractionator in accordance with an embodiment.

[0010] FIG. 3 depicts a field flow fractionator in accordance with an exemplary embodiment.

[0011] FIG. 4 depicts a field flow fractionator in accordance with an exemplary embodiment.

[0012] FIG. 5 depicts a field flow fractionator in accordance with an embodiment. [0013] FIG. 6 depicts a field flow fractionator in accordance with an embodiment.

[0014] FIG. 7 depicts a field flow fractionator in accordance with an embodiment.

DETAILED DESCRIPTION

[0015] The present disclosure describes a field flow fractionator. In an exemplary embodiment, the field flow fractionator includes a spacer including a core, and a coating covering the core. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate

(BoPET). In an embodiment, the plastic is polycarbonate. In an embodiment, the core includes stainless steel. In a particular embodiment, the core is stainless steel. In an embodiment, the coating is silicone. In an embodiment, the coating is polytetrafluoroethylene (PTFE). In an embodiment, the coating is ethylene tetrafluoroethylene (ETFE). In an alternative embodiment, the coating covers only the top side of the core. In an alternative embodiment, the coating covers only the bottom side of the core.

[0016] In an exemplary embodiment, the field flow fractionator includes (1) a spacer including a core, and a coating covering the core, (2) a bottom block including pins, and (3) atop block including mating holes to receive the pins, where the spacer is configured to be positioned between the top block and the bottom block, where the spacer includes mating holes to receive the pins. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET).

In an embodiment, the plastic is polycarbonate. In an embodiment, the coating is polytetrafluoroethylene (PTFE).

[0017] In an exemplary embodiment, the field flow fractionator includes (1) a spacer including (a) a core including an opening, atop side, and a bottom side, (b) a first coating covering the top side of the core, and (c) a second coating covering the bottom side of the core, (2) a top block configured to press on the first coating along a first periphery of the opening, and

(3) a bottom block configured to press on the second coating along a second periphery of the opening. In an embodiment, the core includes plastic. In a particular embodiment, the core is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET).

In an embodiment, the plastic is polycarbonate. In an embodiment, the first coating is polytetrafluoroethylene (PTFE). In an embodiment, the second coating is polytetrafluoroethylene (PTFE).

[0018] In an embodiment, the spacer is soft enough to form a robust seal, while being simultaneously rigid enough to avoid extrusion and to set a reproducible channel thickness. For example, the spacer could be a laminated composite with a rigid core and a soft coating to facilitate the seal.

[0019] In an alternate embodiment, a composite seal such as glass filled PTFE could be used

(e.g., Gylon® 3500 series). The embedded glass microspheres in this material could resist compression while maintaining a good seal.

[0020] Even if the seal were maintained while pressurized, the internal pressure could generate a lateral force on the spacer, which could cause the spacer to extrude and compromise the cell geometry. In order to combat lateral extrusion, in a embodiment, a plurality of pins in the top block along with mating holes in the spacer and bottom block could be used to prevent extrusion. Alternatively, in an embodiment, the pins could be put in the bottom block with mating holes in the upper block. The number of pins required to hold the spacer in place could depend on the rigidity of the spacer material and the ultimate pressure at which the FFF system would operate.

Definitions

Particle [0021] A particle may be a constituent of a liquid sample aliquot. Such particles may be molecules of varying types and sizes, nanoparticles, virus like particles, liposomes, emulsions, bacteria, and colloids. These particles may range in size on the order of nanometer to microns.

Analysis of Macromolecular or Particle Species in Solution

[0022] The analysis of macromolecular or particle species in solution may be achieved by preparing a sample in an appropriate solvent and then injecting an aliquot thereof into a separation system such as a liquid chromatography (LC) column or field flow fractionation

(FFF) channel where the different species of particles contained within the sample are separated into their various constituencies. Once separated, generally based on size, mass, or column affinity, the samples may be subjected to analysis by means of light scattering, refractive index, ultraviolet absorption, electrophoretic mobility, and viscometric response.

Field Flow Fractionation

[0023] The separation of particles in a solution by means of field flow fractionation, FFF, was studied and developed extensively by J. C. Giddings beginning in the early 1960s. The basis of these techniques lies in the interaction of a channel-constrained sample and an impressed field applied perpendicular to the direction of flow. Among those techniques of current interest is cross flow FFF, often called symmetric flow (SF1FFF), where an impressed field is achieved by introducing a secondary flow perpendicular to the sample borne fluid within the channel. There are several variations of this technique including asymmetric flow FFF (i.e., A4F), and hollow fiber (H4F) flow separation.

[0024] Other FFF techniques include (i) sedimentation FFF (SdFFF), where a gravitational/centrifugal cross force is applied perpendicular to the direction of the channel flow,

(ii) electrical FFF (EFFF), where an electric field is applied perpendicular to the channel flow, and (ii) thermal FFF (ThFFF), where a temperature gradient is transversely applied. [0025] Common to all these methods of field flow fractionation is a fluid, or mobile phase, into which is injected an aliquot of a sample whose separation into its constituent fractions is achieved by the application of a cross field. Many of the field flow fractionators allow for the control and variation of the strength of the cross field during the time the sample aliquot flows down the channel, be it electrical field, cross flow, thermal gradient or other variable field.

Symmetric Flow Cross Flow Fractionator (SF1FFF)

[0026] As an illustration of the separation of particles by field flow fractionation, a simplification of perhaps the most straightforward system, a SF1FFF, is described. A sample is injected into an inlet port along with the spending mobile phase. The sample is allowed to undergo a so-called “relaxation phase,” where there is no applied channel flow, but larger particles are forced further down the height of the channel than smaller particles by the constantly applied cross flow. Once the channel flow is resumed, the sample aliquot begins to undergo non-steric separation while it moves down the length channel with the smaller particles leading the larger ones, as they inhabit a region of the cross section of the channel flow nearer the center of the height of the channel where the channel flow is most swift By increasing the cross flow rate, the separation of all species continues while the larger fractions begin to trail further behind their smaller sized companions. After exiting the channel through the outlet port the fractionated sample may be analyzed using various detectors.

Asymmetric Flow FFF ( A4F)

[0027] An asymmetric flow FFF (A4F) is generally considered a variation of the earlier developed SF1FFF. The elements of an A4F channel assembly are depicted in FIG. 1. An A4F channel assembly may include (1) a bottom assembly structure 150 holding a liquid-permeable frit 107 surrounded by a sealing O-ring 105, (2) a permeable membrane that lies on frit 107, (3) a spacer 110 of thickness from about 75 μm to 800 μm into which has been cut a cavity, and (4) a top assembly structure generally holding a transparent plate of polycarbonate material or glass. [0028] The resulting sandwich is held together with bolts or other means, such as applied pressure adequate to keep the channel sealed against leaks, where such pressure may be applied by vise or clamping mechanism so long as it is able to provide relatively even pressure across the channel assembly such that no leaks occur. The generally coffin-shaped or tapered cavity in spacer 110 serves as the channel in which separation will occur. The top assembly structure 140 usually contains three holes, called ports, that pass through the top plate 110 and are centered above the channel permitting the attachment of fittings thereto. These ports are (a) a mobile phase inlet port located near the beginning of the channel and through which is pumped the carrier liquid, the so-called mobile phase, (b) a sample port, downstream of the inlet port, into which an aliquot of the sample to be separated is introduced to the channel and focused thereunder, and (c) an exit port through which the fractionated aliquot leaves the channel near the end of the cavity.

Current Technologies

[0029] FIG. 1 depicts an existing field flow fractionator (FFF) including atop block 140, a spacer 110, an O-ring 105, a frit 107, and a bottom block 150. A field flow fractionator may include atop block, a spacer, and a bottom block, where the bottom block includes an O-ring and a frit For example, the field flow fractionator depicted in FIG. 1 may be an asymmetric field flow fractionator (AF4). An AF4 assembly consists of number of number of parts as shown in

FIG. 1. There is atop plate/top block 140, a spacer 110, an O-ring 105, an ultrafiltration membrane, and a bottom assembly/bottom block 150.

[0030] The spacer serves a number of roles, which require conflicting material properties. It sets the height of the separation channel that controls, along with the analysis method, the fractionation size range, resolution, and sample dilution. It also serves as a seal that prevents fluid from escaping the channel except through the inlet and outlet ports. Lastly, it sets the lateral dimensions of the separation channel. The spacer, when used to define the channel geometry must have a well-defined and reproducible thickness. It also needs to be chemically inert

[0031] As such, common material choices for the spacer are hard materials such as biaxially- oriented polyethylene terephthalate (BoPET) for spacers between 0.1mm and 0.49mm. For spacers from 0.49mm to 3mm, 316 stainless steel sheet is commonly used. Unfortunately these materials are not conformable and tend to make poor seals unless very high clamping forces are used. Even with such high clamping forces, the upper pressure limit for which they would maintain a seal is often less than 30bar. If a soft sealing material were used to improve the seal integrity and reduce the clamping forces required, the seal gasket could extrude horizontally and could compromise the structural integrity of the channel. Thus, there is a need to seal a field flow fractionator via spacers, coatings, and blocks that prevent such extrusion and that preserve the structural integrity of the channel.

[0032] Referring to FIG. 2A, in an exemplary embodiment, the field flow fractionator includes a spacer 210 including a core 220, and a coating 230 covering core 220. In an embodiment, core 220 includes plastic. In a particular embodiment, core 220 is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET). In an embodiment, the plastic is polycarbonate. In an embodiment, core 220 includes stainless steel.

In a particular embodiment, core 220 is stainless steel. In an embodiment, coating 230 is silicone. In an embodiment, coating 230 is polytetrafluoroethylene (PTFE). In an embodiment, coating 230 is ethylene tetrafluoroethylene (ETFE). In an alternative embodiment, coating 230 covers only the top side of core 220. In an alternative embodiment, coating 230 covers only the bottom side of core 220.

TOP Block with Pins

[0033] In further embodiment, the field flow fractionator further includes (a) a top block including pins, and (b) a bottom block including mating holes to receive the pins, where the spacer is configured to be positioned between the top block and the bottom block, where the spacer includes mating holes to receive the pins. Referring to FIG. 2B, in further embodiment, the field flow fractionator further includes (a) a top block 240 including pins 242, and (b) a bottom block 250 including mating holes 252 to receive pins 242, where spacer 210 is configured to be positioned between top block 240 and bottom block 250, where spacer 210 includes mating holes 212 to receive pins 242.

Bottom Block with Pins

[0034] In further embodiment, the field flow fractionator further includes (a) a bottom block including pins, and (b) a top block including mating holes to receive the pins, where the spacer is configured to be positioned between the top block and the bottom block, where the spacer includes mating holes to receive the pins. Referring to FIG. 2C, in further embodiment, the field flow fractionator further includes (a) a bottom block 260 including pins 262, and (b) a top block

270 including mating holes 272 to receive pins 262, where spacer 210 is configured to be positioned between top block 270 and bottom block 260, where spacer 210 includes mating holes

212 to receive pins 262.

Spacer with Blocks. Pins, and Holes

[0035] Referring to FIG. 3, in an exemplary embodiment, the field flow fractionator includes flow fractionator includes (1) spacer 210 including core 220, and a coating 230 covering core

220, (2) a bottom block 360 including pins 362, and (3) atop block 370 including mating holes

372 to receive pins 362, where spacer 210 is configured to be positioned between top block 370 and bottom block 360, where spacer 210 includes mating holes 212 to receive pins 362. In an embodiment, core 220 includes plastic. In a particular embodiment, core 220 is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (Bo PET). In an embodiment, the plastic is polycarbonate. In an embodiment, coating 230 is polytetrafluoroethylene (PTFE). Spacer with Blocks

[0036] Referring to FIG. 4, in an exemplary embodiment, the field flow fractionator includes

(1) a spacer 410 including (a) a core 420 including an opening 422, atop side 426, and a bottom side 428, (b) a first coating 430 covering top side 426 of core 420, and (c) a second coating 435 covering bottom side 428 of core 420, (2) a top block 440 configured to press on first coating

430 along a first periphery 423 of opening 422, and (3) a bottom block 450 configured to press on second coating 435 along a second periphery 425 of opening 422. In an embodiment, core

420 includes plastic. In a particular embodiment, core 420 is plastic. In an embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET). In an embodiment, the plastic is polycarbonate. In an embodiment, first coating 430 is polytetrafluoroethylene (PTFE). In an embodiment, second coating 435 is polytetrafluoroethylene (PTFE).

Alternatives

[003η In an embodiment, as depicted in FIG. 5, the field flow fractionator includes (1) a spacer including (a) a core including an opening, (b) a first coating covering the core along a first periphery of the opening, and (c) a second coating covering the core along a second periphery of the opening, (2) a top block configured to press on the first coating, and (3) a bottom block configured to press on the second coating. In an embodiment, the core includes stainless steel. In an embodiment, the core is stainless steel. In an embodiment, the core includes plastic. In an embodiment, the core is plastic. In a particular embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET). In an embodiment, the first coating is silicone. In an embodiment, the first coating is polytetrafluoroethylene (PTFE). In an embodiment, the first coating is ethylene tetrafluoroethylene (ETFE). In an embodiment, the second coating is silicone. In an embodiment, the second coating is polytetrafluoroethylene

(PTFE). In an embodiment, the second coating is ethylene tetrafluoroethylene (ETFE). In an alternative embodiment, the field flow fractionator includes (1) a spacer including (a) a core including an opening, (b) a coating covering the core along a periphery of the opening, and (2) a top block configured to press on the coating. In an alternative embodiment, the field flow fractionator includes (1) a spacer including (a) a core including an opening, and (b) a coating covering the core along a periphery of the opening, and (2) a bottom block configured to press on the coating.

[0038] In an embodiment, as depicted in FIG. 6, the field flow fractionator includes (1) a spacer including an opening, (2) atop block configured to press on the spacer along a first periphery of the opening, and (3) a bottom block configured to press on the spacer along a second periphery of the opening. In an embodiment, the spacer includes stainless steel. In an embodiment, the spacer is stainless steel. In an embodiment, the spacer includes plastic. In an embodiment, the spacer is plastic. In a particular embodiment, the plastic is biaxially-oriented polyethylene terephthalate (BoPET).

[0039] In an exemplary embodiment, as depicted in FIG. 7, the field flow fractionator includes (1) a spacer including an opening, a top side, and a bottom side, and (2) a top block including a bottom side and a coating covering the bottom side of the top block, resulting in a coated bottom side of the top block, where the coated bottom side of the top block is configured to press on the top side of the spacer along a periphery of the opening. In an embodiment, the spacer includes stainless steel. In an embodiment, the spacer is stainless steel. In an embodiment, the spacer includes plastic. In an embodiment, the spacer is plastic. In a particular embodiment, the plastic is biaxially-oriented polyethylene terephthalate (Bo PET). In an embodiment, the coating is silicone. In an embodiment, the coating is polytetrafluoroethylene

(PTFE). In an embodiment, the coating is ethylene tetrafluoroethylene (ETFE). In an alternative embodiment, the field flow fractionator includes (1) spacer including an core including an opening, a top side, and a bottom side, and (b) a spacer coating covering the top side of the core, and (2) a top block including a bottom side and a top block coating covering the bottom side of the top block, resulting in a coated bottom side of the top block, where the coated bottom side of the top block is configured to press on the top side of the spacer along a periphery of the opening. In an alternative embodiment, the field flow fractionator includes (1) spacer including an core including an opening, a top side, and a bottom side, and (b) a spacer coating covering the bottom side of the core, and (2) a top block including a bottom side and a top block coating covering the bottom side of the top block, resulting in a coated bottom side of the top block, where the coated bottom side of the top block is configured to press on the top side of the spacer along a periphery of the opening. In an alternative embodiment, the field flow fractionator includes (1) spacer including an core including an opening, a top side, and a bottom side, (b) a first spacer coating covering the top side of the core, and (c) a second spacer coating covering the bottom side of the core, and (2) a top block including a bottom side and a top block coating covering the bottom side of the top block, resulting in a coated bottom side of the top block, where the coated bottom side of the top block is configured to press on the top side of the spacer along a periphery of the opening.

Example

[0040] For example, spacers made of biaxially-oriented polyethylene terephthalate (BoPET) with silicone coating (e.g., DOWSIL™ 1-2755), spacers made of 316 Stainless with silicone, and spacers made of stainless steel with PTFE coating were tested. All such spacers were tested by filling the cell assembly with water, capping the ports except for one, and pressurizing the channel through the open port to 30bar. The FFF system was sealed, and the pressure was monitored over time. The rate of pressure loss was a measure of the leak rate. Success was defined as holding the applied pressure with 2% over a span of 30 minutes. All non-composite spacer configurations that were tested failed this test within 5 minutes. The spacers that had been coated with PTFE or with silicone (e.g., Dow-Sil) passed this test [0041] Also, the spacers made of BoPET could require more pins than spacers made of stainless steel. The spacers made of Gylon could seal well, but could require a large number of restraining pins or a restraining flange to resist extrusion.

[0042] The descriptions of the various embodiments of the present disclosure have been presented for purposes of illustration, but are not intended to be exhaustive or limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terminology used herein was chosen to explain the principles of the embodiments, the practical application or technical improvement over technologies found in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.