Related Application

This application claims priority to U.S. provisional application no. 62/984,192 filed on March 2, 2020 the content of which is incorporated herein by reference in its entirety.

Background

The present teachings are generally related to an electromagnetic device that can be employed in magnetic particle based sampling techniques.

Vacutainers and other similar tubes/vials with rubber caps are widely used in various fields, and particularly in clinical laboratories. Samples can be loaded and accessed in such containers through a needle piercing through the rubber cap without opening the container, thus reducing potential biohazard, sample contamination, and solvent evaporation.

Solid-phase microextraction (SPME) devices have been developed, which can perform sampling, sample preparation and extraction in one step, thus greatly simplifying sample analysis. Such SPME devices can include a fiber that is protected within a needle, which can be used to pierce a septum of a sealed container. Once the needle is inserted into the container, the fiber can be pushed further down to be immersed into the sample for extraction of one or more analytes of interest. After the extraction process, the fiber is pulled back into the protection needle and is removed from the sample vial.

Although SPME devices have been proved to be successful in extraction and transfer of samples, they exhibit certain shortcomings. For example, the binding capacity of a conventional SPME fiber is limited by its small surface area. Although significant enhancement of extraction efficiency can be obtained when an SPME membrane having a larger surface area is employed, the integration of an SPME membrane with a rubber piercing needle is difficult.

Summary

An electromagnetic sampling device is disclosed, which comprises a needle having a hollow housing that extends from a proximal end to a distal end, and an electromagnet comprising an electromagnetic coil and a metal core, at least a portion of said metal core extending through said hollow housing of the needle and be configured to transition between an extended position in which the distal end of the metal core extends beyond the distal end of the needle's hollow housing and a retracted position in which the distal end of the metal core is positioned within the needle's housing. The activation of the electromagnetic coil can magnetize the metal core. Such activation can be achieved in the retracted and extended positions.

The electromagnetic sampling device further comprises a flange that is coupled to said metal core for moving the metal core between the extended and retracted positions. In some embodiments, the needle's housing is configured at its distal end for piercing a septum employed to seal a container.

In some embodiments, the electromagnet is positioned outside of the needle's hollow housing.

The metal core can be configured to collect a plurality of magnetic particles disposed in the container when the metal core is magnetized via activation of the electromagnetic coil. In some embodiments, the hollow housing of the needle is substantially cylindrical. By way of example, in some embodiments, such a hollow cylindrical housing can have an inner diameter equal to or greater than about 0.5 mm. By way of example, the inner diameter of the hollow cylindrical housing can be in range of about 0.5 mm to about 10 mm.

In some embodiments, the needle's housing comprises a magnetic shielding material, such as a ferromagnetic metal or Mu Metal. In some embodiments, the metal core can be formed of any of silicon steel, or ferrite.

In some embodiments, in an extended position, the distal end of the metal core extends beyond the distal end of the needle's housing for a length in a range of about 1 mm to about 100 mm.

In a related aspect, a method of collecting magnetic particles from a particle container and transferring the collected magnetic particles to a container sealed by a septum using an electromagnetic sampling device according to the present teachings is disclosed. Such a method can include inserting at least the distal end of the needle within a container containing a plurality of magnetic particles. Typically, the distal end of the needle is inserted into the container with the metal core in a retracted position. Subsequently, the metal core can be transitioned from the retracted position to the extended position such that the distal end of the metal core will be in proximity of the magnetic particles. The electromagnetic coil can be activated so as to magnetize the metal core and at least some of the magnetic particles can be captured via the magnetized metal core. The metal core and the associated collected magnetic particles can then be transitioned from the extended position into the retracted position and the needle and the captured metal particles can be removed from the particle container.

The needle can then be used to pierce a septum of a sealed container in which one or more target analytes are disposed and at least the distal end of the needle can be inserted into the sealed container. The metal core can be transitioned from the retracted position into the extended position and the electromagnet can be deactivated so as to release the captured magnetic particles into the container in which the target analyte(s) are disposed. In some embodiments, the magnetic particles are functionalized, e.g., coated, so as to capture the target analytes. By way of example, in some embodiments, the magnetic particles can be functionalized with antibodies that exhibit specific binding to the target analytes. In other embodiments, the magnetic particles can be functionalized with C18, antibody or any other suitable moiety.

In some embodiments, the mixing of the magnetic particles is performed so as to facilitate the capture of the target analytes by those particles. Such mixing can be achieved, for example, via a AC mixing device. In some embodiments, an RF source can be coupled to the electromagnetic sampling device to apply an AC signal to its metal core so as to facilitate 3-dimensional (3D) mixing of the magnetic particles. In some such embodiments, the AC signal can have a frequency in a range of about 1 Hz to about 400 Hz. The field strength generated by the AC signal can be, for example, in a range between 50 to 200 mT, e.g., in a range of about 20 to 100 mT.

The metal core can then be retracted to bring the magnetic particles together with the target analytes attached thereto into the needle's housing. The needle can then be removed from the target container to extract the magnetic particles and the associated attached target analytes.

In some embodiments, the distal end of the needle can be inserted into an open port sampling interface of a mass spectrometer and the magnetic particles can be released into the mass spectrometer, via transitioning the metal core from a retracted position into an extended position and deactivating the electromagnet. In some embodiments, the magnetic particles to which analytes are attached are washed prior to introducing them into the open port sampling interface.

In some embodiments, the electromagnet is kept energized as a continuous liquid flow in the OPP washes out analytes from the magnetic particles (beads). In some embodiments, the magnetic particles can be released into the OPP interface and an electromagnetic mixer can be used to mix the released particles in the OPP sampling port. By way of example, the teachings of U.S. Published Application 2018/0369831, which is herein incorporated by reference in its entirety, can be employed for such mixing of the magnetic particles. Alternatively, the teachings of U.S. Published Application 2020/0043712, which is also herein incorporated by reference in its entirety, can be employed to provide such mixing of the magnetic particles. Alternatively, in some embodiments, the magnetic particles can enter the OPP interface and they travel to the drain or can be trapped/collected somewhere downstream.

Further understanding of various aspects of the present teachings can be obtained by reference to the following detailed description in conjunction with the associated drawings, which are described briefly below.

Brief Description of the Drawings

FIGs. 1A and IB schematically depict an electromagnetic device according to an embodiment of the present teachings,

FIG. 2 depicts an electromagnetic device according to the present teachings inserted into a container that contains a plurality of magnetic particles,

FIG. 3A depicts the electromagnetic device of FIG. 2 with the magnetic core together with magnetic particles attached thereto in a retracted position, where the distal end of the needle is employed for piercing a septum of a sealed container in which one or more target analytes are disposed,

FIG. 3B shows the electromagnetic device of FIG. 3A with the magnetic core and the attached magnetic particles retracted into the needle's housing,

FIG. 4 is a flow chart depicting various steps in a method according to an embodiment for extracting magnetic particles from a container and transferring the extracted magnetic particles to another container,

FIG. 5 is a schematic view of a fluid mixing system, which can be employed to agitate and mix a plurality of magnetic particles.

Detailed Description

The present teachings are generally directed to an electromagnetic sampling device, which includes a protective needle and an electromagnet comprising an electromagnetic coil and a magnetizable metal core that extends at least partially through the hollow housing of the protective needle. The metal core can be transitioned between an extended and a retracted position to allow transferring magnetic particles from one container to another. In some embodiments, the electromagnetic sampling device can be employed to introduce analytes attached to a plurality of magnetic particles into an input port of a mass spectrometer. FIGs. 1A and IB schematically depict an electromagnetic sampling device 100 according to an embodiment, which includes a needle 102 characterized by a hollow cylindrical housing, which extends from a proximal end (PE) to a distal end (DE). In some embodiments, the distal end of the needle is configured for piercing a septum of a sealed container, as discussed in more detail below.

The electromagnetic sampling device 100 further includes an electromagnet 104 that has an electromagnetic coil 106 and a magnetizable metal core 108 that can be magnetized by the electromagnetic coil, when the coil is activated. The metal core 108 extends through the hollow enclosure of the needle's housing from its proximal end to its distal end. A flange 110 attached to the metal core allows transitioning the magnetizable metal core from a retracted position in which a distal end of the metal core is within the needle's housing (FIG. 1A) into an extended position in which the distal end of the metal core extends beyond the distal end of the needle's housing (FIG. IB). As discussed in more detail below, once magnetized, in the extended position, the metal core can collect magnetic particles disposed within a particle container, which can then be transferred to another container containing one or more analytes of interest. In some embodiments, the metal core is transitioned by a mechanical actuator. In some embodiments, the activation of the electromagnetic coil can be linked to the location of the metal core. For example, in one embodiment, the electromagnetic coil is energized when the metal core is in an extended position and the electromagnetic coil is de-energized when the metal core is in a retracted position.

As discussed in more detail below, in many embodiments, the magnetic particles are functionalized, e.g., coated, with one or more moieties that can specifically bind to one or more analytes to allow their extraction, e.g., from a container. Further, in some embodiments, the target analytes extracted via the magnetic particles can be introduced into an open port interface of a mass spectrometer.

As discussed in more detail below, the needle's distal end is shaped and sized so as to allow its penetration through a septum of a sealed container. As noted above, in this embodiment, the needle 102 has a hollow housing that is substantially cylindrical and has an inner diameter equal to or greater than about 0.5 mm. By way of example, in some embodiments, the inner diameter of the needle's housing can be in a range of about 0.5 mm to about 10 mm. In general, the inner diameter of the needle's housing is selected to ensure that it can accommodate the metal core (and magnetic particles collected by the metal core as discussed more below) and further allow using the needle to pierce through a septum sealing a container. The needle can have a variety of different lengths. By way of example, the length of the needle (i.e., the distance between its proximal and distal ends) can be in a range of about 0.5 cm to about 20 cm. In general, the diameter of the portion of the metal core intended to be received within the needle's housing is selected so as to allow its insertion into the needle's housing with sufficient clearance relative to the needle's inner wall such that magnetic particles collected by the distal end of the needle can be brought into the needle's housing when the metal core is transitioned from the extended position into the retracted position. By way of example, in some embodiments, the metal core (or at least a portion thereof that is intended for insertion into the needle's housing), can have a diameter in a range of about 0.1 mm to about 10 mm, e.g., in a range of about 0.3 mm to about 5 mm. The length of the portion of the metal core that extends beyond the distal end of the needle's housing in the extended position can be, for example, in a range of about 1 mm to about 10 mm.

The needle can be formed of a variety of different materials. In this embodiment, the needle is formed of a magnetic shielding material. The use of a magnetic shielding material for forming needle can advantageously inhibit, and preferably prevent, the magnetic particles from being trapped on the external surface of the needle. Some suitable examples of such magnetic shielding materials include, without limitation, a ferromagnetic metal or MuMetal (a nickel-iron soft ferromagnetic alloy having a high permeability).

In this embodiment, the electromagnetic coil 106 is positioned outside the needle. In some embodiments, the electromagnetic coil assembly can have an outer diameter, for example, in a range of about 0.1 to about 100 mm, e.g., about 20 mm.

With continued reference to FIGs. 1A, IB as well as FIGs. 2, 3A and 3B and the flow chart of FIG. 4, the above electromagnetic sampling device 100 can be used to collect magnetic particles (herein also referred to as magnetic beads) contained in a particle container and transfer those particles to another container in which one or more analytes of interest are contained. The magnetic particles can be formed, e.g., from a magnetic metal, such as a magnetic alloy. By way of example, in some embodiments, the magnetic particles can have a silica core with a metal coating, or can have metal cores. In some embodiments, the outer surfaces of the magnetic particles can be functionalized, e.g., with an antibody or C18, to allow the magnetic particles to bind to one or more analytes of interest.

For example, in one method for transferring magnetic particles from one container to another using an electromagnetic sampling device according to the present teachings, at least the distal end of the needle 102 is inserted into a particle container 200 in which a plurality of magnetic particles 202 are disposed. As indicated above, the magnetic particles can be functionalized to collect one or more analytes of interest. Typically, such insertion of the needle into the particle container is done with the magnetizable metal core 108 in a retracted position within the needle's housing, though in other cases, the needle can be inserted into the particle container with the magnetizable metal core in an extended position. The magnetizable metal core 108 can then be transitioned from its retracted position into its extended position using the flange 110 such that the distal end of metal core is in the vicinity of the magnetic particles, as shown in FIG. 2. The electromagnetic coil 106 can be activated (either before or after transitioning the metal core from a retracted position to an extended position) so as to magnetize the metal core. The distal end of the magnetized metal core can then attract the magnetic particles so as to collect at least a portion of those particles, as shown schematically in 2.

The distal end of the metal core and the magnetic particles attached thereto can then be retracted into the needle's protective housing and the needle can be removed from the particle container. The electromagnetic coil remains energized to ensure that the magnetic particles continue to remain attached to the magnetized distal end of the metal core. In many embodiments, the maximum of the magnetic field strength is at the tip of the metal core, where typically most of the magnetic particles are trapped. To quantitatively aliquot the magnetic particles out, in some embodiments, the magnetic particles are suspended in the solution and can be pre-agitated (e.g., via mechanical shaking) or with the electromagnetic mixer.

The distal end of the needle can then be employed to pierce a septum 302 sealing a container 300 in which a sample 304 containing at least one analyte of interest 305 is disposed, as shown schematically in FIG. 3A. For example, the analyte can be dispersed within a medium 306, such as a liquid. Once inserted into the sealed container 300, the metal core 108 can be transitioned from the retracted position into the extended position so as to bring the distal end of the metal core in vicinity of the analyte 304, as shown in FIG. 3B. The electromagnetic coil can then be deactivated so as to demagnetize the metal core, thereby releasing the captured magnetic particles into the container 300.

In some embodiments, the released functionalized magnetic particles can be manipulated to facilitate the capture of the target analyte(s) in the sample. By way of example, AC mixing of the functionalized released particles can be used to facilitate capture of the target analyte(s) by the released particles. For example, published PCT Application No. PCT/IB2018/050399 entitled "Electromagnetic Assemblies for Processing Fluid," which is herein incorporated by reference in its entirety, discloses methods and systems for mixing fluids, which can be employed in some embodiments of the present teachings. This publication generally discloses a fluid processing system that includes a magnetic assembly having a plurality of magnetic structures that are configured to generate a magnetic field gradient within a fluid container. The magnetic structures may be formed as a plurality of electromagnets that may be individually actuated by a controller, where each electromagnet can generate a magnetic field within the container. More specifically,

FIG. 2 of this publication, which is reproduced herein as FIG. 5, depicts such a fluid processing magnetic assembly 204 that includes an upper magnetic structure 245a and a lower magnetic structure 245b, each of which includes four electromagnets 210 comprising an electrically- conductive wire 212 in the form of a coil. The inner ends of the electromagnets 210a-d are spaced apart from the central axis so as to receive a fluid container therebetween. An AC signal can be applied to the various electromagnets 210a-d such that the magnetic field gradients change over time, thereby causing the fluid to experience mixing due to the corresponding movement of magnetic particles within the fluid container.

Further, in some embodiments, as shown schematically in FIG. 3B, an RF source 400 can be coupled to the metal core 108 to apply an RF signal thereto in order to enhance the 3-D mixing of the magnetic particles, e.g., via mixing the medium in which the analyte is dispersed. The mixing of the particles can advantageously enhance their ability to capture the analyte(s) of interest.

Subsequent to capturing the analyte(s) of interest, the electromagnetic sampling device 100 can be used to remove the magnetic particles from the sealed container. In particular, the electromagnetic coil 106 can be re-energized to magnetize the metal core and the magnetized metal core can be employed to capture the magnetic particles, e.g., by trapping the particles at the distal end of the metal core. The metal core together with the captured magnetic particles can be retracted into the protective needle's housing and the needle can be removed from the sealed container.

In some embodiments, the magnetic particles can include at least two sets of particles that are functionalized differently so as to capture different types of analytes. For example, in some such embodiments, one set of magnetic particles can be functionalized so as to capture proteins and another set of magnetic particles can be functionalized so as to capture lipids. In such cases, sequential extraction of different analytes within a sample using different magnetic particles can be performed independently using the methods discussed above. In such embodiments, the extraction of one analyte does not adversely affect the extraction of another analyte.

In some embodiments, functionalized magnetic particles can be employed for pre-treatment of an analyte of interest. By way of example, trypsin-coated magnetic particles can be introduced into a sample for digestion of one or more analytes of interest. The trypsin-coated magnetic particles can then be removed and C18 functionalized magnetic particles can be introduced into the sample, e.g., to extract peptides.

In some embodiments, the sampling device can be mechanically connected to a manipulator. The manipulator can be configured to operatively position the sampling device above a sample such that the distal end extends to contact a sample in a first container when in the extended position and retracts the distal end away from the sample when in a retracted position. In some embodiments, the manipulator can include a robotic arm. In some embodiments, the manipulator can be further configured to and be operative to locate the sampling device above a second container after capturing the magnetic particles from a first container. The second container can contain a liquid (e.g., solvent) that can be utilized to wash the distal end and any magnetic particles contained therein. In some embodiments, the manipulator is further configured to and is operative to position the sampling device opposite the open end of an open port sampling interface such that when in the extended position, the distal end extends and is immersed into the solvent flowing at the open end of the open port sampling interface and retracts out of the solvent when retracted into the retracted position.

An electromagnetic sampling device according to the present teachings provides a number of advantages. For example, such a device can provide a higher surface area for capturing magnetic particles, thereby increasing extraction efficiency. Further, such an electromagnetic sampling device can be integrated with RF frequency mixing, as discussed above. Those having ordinary skill in the art will appreciate that various changes can be made to the above embodiments without departing from the scope of the invention.