阅读说明：本技术 磁性辅助分离装置及相关方法 (Magnetically assisted separation device and related methods ) 是由 C.M.舒尔斯 J.J.普罗夫基 J.K.卡乌亚 于 2018-06-27 设计创作，主要内容包括：提供了用于从其中悬浮有目标物质的培养基中分离该目标物质的磁性辅助分离装置以及相关方法。根据一个方面,磁性分离器可以包括具有开口的框架,该开口被配置为接纳包含该培养基的一个或多个容器。此外,该磁性分离器可以包括安装在该框架的相对侧上的第一磁场发生元件和第二磁场发生元件,使得该一个或多个容器可以定位在该第一磁场发生元件与该第二磁场发生元件之间。根据另一个方面,工作站包括：工作表面,该工作表面用于接纳包含该培养基的一个或多个容器；流体转移构件；自动操纵器,该自动操纵器被配置为使该流体转移构件移动；以及多个磁场发生元件,每个磁场发生元件能够在远离该一个或多个容器的第一位置与邻近该一个或多个容器的第二位置之间移动。(Magnetically assisted separation devices and related methods are provided for separating a target substance from a medium in which the target substance is suspended. According to one aspect, a magnetic separator may include a frame having an opening configured to receive one or more vessels containing the culture medium. Furthermore, the magnetic separator may comprise a first magnetic field generating element and a second magnetic field generating element mounted on opposite sides of the frame such that the one or more containers may be positioned between the first magnetic field generating element and the second magnetic field generating element. According to another aspect, a workstation comprises: a working surface for receiving one or more containers containing the medium; a fluid transfer member; a robotic manipulator configured to move the fluid transfer member; and a plurality of magnetic field generating elements, each magnetic field generating element being movable between a first position distal from the one or more receptacles and a second position proximal to the one or more receptacles.)

1. A magnetic separator for separating a target substance from a culture medium having the target substance suspended therein, the culture medium contained in at least one container, the magnetic separator device comprising:

a frame having a first opening configured to receive the at least one container; and

a first magnetic field generating element and a second magnetic field generating element mounted on opposite sides of the frame at a distance from each other such that the at least one container can be positioned between the first magnetic field generating element and the second magnetic field generating element.

2. The magnetic separator of claim 1 wherein the frame includes a first vertical wall and a second vertical wall, the first magnetic field generating element being mounted on an inwardly facing surface of the first vertical wall and the second magnetic field generating element being mounted on an inwardly facing surface of the second vertical wall.

3. The magnetic separator of claim 2, the first vertical wall being parallel to the second vertical wall.

4. The magnetic separator of any of claims 2 or 3 including at least one of a beam or a third wall extending between the inwardly facing surface of the first vertical wall and the inwardly facing surface of the second vertical wall.

5. The magnetic separator of any of claims 2-4, the first opening being defined between the first vertical wall and the second vertical wall and allowing the at least one container to be inserted into the magnetic separator in a horizontal direction.

6. The magnetic separator of claim 5 comprising a second opening formed in the frame and allowing access to the at least one receptacle from above when the at least one receptacle is positioned between the first and second magnetic field generating elements.

7. The magnetic separator of any of claims 2 to 6, comprising a non-magnetic support frame configured to hold the at least one receptacle and sized to be inserted through the first opening to position the at least one receptacle between the first and second magnetic field generating elements.

8. The magnetic separator of claim 7, the first end of the non-magnetic support frame fitting below the first magnetic field generating element and the second end of the non-magnetic support frame fitting below the second magnetic field generating element when inserted through the first opening.

9. The magnetic separator of any of claims 7 or 8 in which the at least one receptacle comprises a plurality of receptacles, the non-magnetic support frame being configured to prevent lateral movement of the plurality of receptacles relative to one another.

10. A magnetic separator as claimed in any of claims 1 to 9, in which the first magnetic field generating element comprises a first permanent magnet.

11. A magnetic separator as claimed in claim 10, the first permanent magnet having a maximum magnetic pull equal to or greater than 100 newtons (N).

12. A purification method comprising:

adding a medium in which a target substance is suspended to a vessel;

adding a plurality of magnetic beads to the container, the target substance temporarily binding to the plurality of magnetic beads; and

positioning the container between a first magnetic field generating element and a second magnetic field generating element, the first magnetic field generating element and the second magnetic field generating element being held at a distance from each other by a frame, at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracting and holding the plurality of magnetic beads against an inner surface of the container.

13. The purification method of claim 12, wherein positioning the container between the first and second magnetic field generating elements comprises manually moving the container from a first position external to the frame to a second position between the first and second magnetic field generating elements.

14. The purification method of any one of claims 12 or 13, comprising removing the culture medium from the vessel while at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracts and holds the plurality of magnetic beads against an inner surface of the vessel.

15. The purification method of claim 14, comprising, after removing the culture medium from the container, adding an eluent to the container to elute the target species bound to the plurality of magnetic beads, and removing the container from between the first magnetic field generating element and the second magnetic field generating element such that the plurality of magnetic beads are freely dispersed in the eluent.

16. The purification method of claim 15, comprising repositioning the container between the first magnetic field generating element and the second magnetic field generating element such that at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracts and holds the plurality of magnetic beads against an inner surface of the container, followed by removing the eluate from the container.

17. The purification method of claims 14-16, comprising adding a washing fluid to the vessel and removing the vessel from between the first magnetic field generating element and the second magnetic field generating element after removing the culture medium from the vessel such that the plurality of magnetic beads are freely dispersed in the washing fluid.

18. The purification method of claim 17, comprising repositioning the vessel between the first magnetic field generating element and the second magnetic field generating element such that at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracts and holds the plurality of magnetic beads against an inner surface of the vessel, followed by removing the washing fluid from the vessel.

19. The purification method of claims 12-18, the frame comprising a first vertical wall and a second vertical wall, the first magnetic field generating element mounted on an inward facing surface of the first vertical wall, and the second magnetic field generating element mounted on an inward facing surface of the second vertical wall.

20. The purification method of claim 19, wherein positioning the container between the first and second magnetic field generating elements comprises inserting the container horizontally through an opening defined between the first and second vertical walls.

21. The purification method of any one of claims 12 to 20, the first magnetic field generating element comprising a first permanent magnet.

22. The purification method of claim 21, the first permanent magnet having a maximum magnetic pull equal to or greater than 100N.

23. A workstation for separating a target substance from a culture medium having the target substance suspended therein, the culture medium contained in at least one container, the workstation comprising:

a working surface for receiving the at least one container containing the culture medium;

a fluid transfer member configured to transfer fluid to and from the at least one container;

a robotic manipulator configured to move the fluid transfer member relative to the working surface; and

a plurality of magnetic field generating elements, each magnetic field generating element being movable relative to the working surface between a first position distal from the at least one container and a second position adjacent to the at least one container.

24. The workstation of claim 23, the plurality of magnetic field generating elements being laterally spaced from one another to define a plurality of rows for receiving a plurality of containers, the plurality of containers comprising: the at least one container.

25. The workstation of claim 24, each magnetic field generating element of the plurality of magnetic field generating elements having a longitudinal axis, the longitudinal axes of the plurality of magnetic field generating elements being parallel to each other.

26. The workstation of claim 25, each magnetic field generating element of the plurality of magnetic field generating elements having a length parallel to the longitudinal axis and a width perpendicular to the longitudinal axis, wherein the length is greater than or equal to 10 inches and the width is greater than or equal to 0.5 inches.

27. The workstation of any of claims 23 to 26, the work surface being arranged horizontally, each magnetic field generating element of the plurality of magnetic field generating elements being movable in a horizontal direction.

28. The workstation according to any one of claims 23 to 27, comprising a linear actuator configured to reciprocate at least a first of the plurality of magnetic field generating elements between the first and second positions.

29. The workstation as recited in claim 28, the linear actuator including at least one of a hydraulic cylinder, a pneumatic cylinder, or an electric motor.

30. The workstation of any of claims 23 to 29, the robotic manipulator being disposed above the work surface, and the fluid transfer member being configured to transfer fluid to and from the at least one container via an opening on a top of the at least one container.

31. The workstation according to any one of claims 23 to 30, the robotic manipulator comprising a cartesian robot movable in at least an x-direction and a y-direction.

32. The workstation according to any one of claims 23 to 31, comprising a pump in fluid communication with the fluid transfer member and configured to add or remove fluid to or from the at least one container via the fluid transfer member.

33. The workstation of claim 32, the pump mounted on the robotic manipulator.

34. The workstation of any one of claims 32 or 33, comprising a multi-position valve in fluid communication with the pump and configured to selectively couple the pump to at least one of an external fluid source or a drain.

35. The workstation of any one of claims 23 to 34, comprising a plurality of magnetic beads for submersion in the media in the at least one container, each magnetic bead of the plurality of magnetic beads having an outer surface configured to temporarily bind with the target substance to separate the target substance from the media.

36. The workstation as claimed in any one of claims 23 to 13, wherein the fluid transfer member comprises at least two parallel fluid conduits.

37. The workstation according to any one of claims 23 to 36, the first magnetic field generating element comprising a first permanent magnet.

38. The workstation as recited in claim 37, the first permanent magnet having a maximum magnetic pull force equal to or greater than 100N.

39. A purification method comprising:

providing a workstation having a working surface, a robotic manipulator movable relative to the working surface and carrying a fluid transfer member, and a plurality of magnetic field generating elements laterally spaced from one another to define a plurality of rows;

adding at least one culture medium having a target substance suspended therein to a plurality of containers;

adding a plurality of magnetic beads to the plurality of containers, the target substance temporarily binding to the plurality of magnetic beads; and

arranging the plurality of containers in a plurality of rows defined between the plurality of magnetic field generating elements such that the plurality of magnetic field generating elements magnetically attract and hold the plurality of magnetic beads against an inner surface of respective ones of the plurality of containers.

40. The purification method of claim 39, comprising causing the robotic manipulator to insert an end of the fluid transfer member into at least one media contained in at least a first vessel of the plurality of vessels and remove the at least one media from the first vessel via the fluid transfer member.

41. The purification method of claim 40, comprising moving at least a first magnetic field generating element of the plurality of magnetic field generating elements from a first position proximate to the first vessel to a second position distal to the first vessel after removing the culture medium from the first vessel.

42. The purification method of claim 41, comprising adding an eluent to the first container via the fluid transfer member to elute the target species bound to the plurality of magnetic beads.

43. The purification method of claim 42, comprising moving the first magnetic field generating element from the second position to the first position such that the first magnetic field generating element magnetically attracts and holds the plurality of magnetic beads against an inner surface of the first container, followed by removing the eluate from the first container via the fluid transfer member.

44. The purification method of any one of claims 41 to 43, comprising adding a washing fluid to the first vessel via the fluid transfer member.

45. The purification method of claim 44, comprising moving the first magnetic field generating element from the second position to the first position such that the first magnetic field generating element magnetically attracts and holds the plurality of magnetic beads against an inner surface of the first vessel, and removing the washing fluid from the first vessel via the fluid transfer member.

46. The purification method of any one of claims 39 to 45, the plurality of magnetic field generating elements comprising a first permanent magnet.

47. The purification method of claim 46, the first permanent magnet having a maximum magnetic pull equal to or greater than 100N.

48. The purification method of any one of claims 39 to 47, the robotic manipulator comprising a Cartesian coordinate robot movable in at least an x-direction and a y-direction.

Technical Field

The present disclosure relates generally to separation devices and methods, and more particularly to separating a target substance (such as a biomolecule) from a fluid medium having the target substance suspended therein to facilitate downstream processing or analysis of the target substance.

Background

Certain diagnostic, research and drug production activities benefit from or require isolation of target substances (such as proteins) contained in cell cultures or other biological mixtures. This task has been accomplished in the past by various techniques. Some of these techniques require that the solubility of the target substance be altered so that it precipitates out of the biological mixture. Certain other techniques require centrifugation, in which particles of different densities are separated by rotating them at high speed around a fixed point. Still other techniques are based on chromatography, which requires passing the biological mixture through a filter material in which the constituent components of the biological mixture move at different rates.

Such conventional purification techniques and others tend to be time consuming, labor intensive, and/or limited to relatively small sample sizes. For example, centrifugation is typically performed in test tubes or bottles, which may limit the amount of material that can be processed at a given time. Moreover, certain conventional purification techniques may require laboratory technicians to manually pipette fluids between various containers, which may be inefficient and may increase the risk of cross-contamination.

The present disclosure sets forth purification-related devices and methods that implement advantageous alternatives to existing purification devices and methods, and that may address one or more of the challenges or needs noted herein as well as provide other benefits and advantages.

Disclosure of Invention

One aspect of the present disclosure provides a magnetic separator for separating or removing a target substance from a medium in which the target substance is suspended. The magnetic separator may include a frame having a first opening configured to receive at least one container or vessel containing a culture medium. The magnetic separator may further comprise a first magnetic field generating element and a second magnetic field generating element. The first magnetic field generating element and the second magnetic field generating element may be mounted on opposite sides of the frame at a distance from each other such that the at least one container can be positioned between the first magnetic field generating element and the second magnetic field generating element.

Another aspect of the disclosure provides a purification method, which may include: (a) adding a culture medium having a target substance suspended therein to a container or vessel; (b) adding a plurality of magnetic beads to the container, the target substance temporarily binding to the plurality of magnetic beads; and (c) positioning the container between a first magnetic field generating element and a second magnetic field generating element, the first magnetic field generating element and the second magnetic field generating element being held at a distance from each other by a frame, at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracting and holding the plurality of magnetic beads against an inner surface of the container.

Additional aspects of the present disclosure provide a workstation for separating a target substance from a medium in which the target substance is suspended. The workstation may include: a working surface for receiving at least one container or vessel containing the culture medium; a fluid transfer member configured to transfer fluid to and from the at least one container. Further, the workstation may comprise: a robotic manipulator configured to move the fluid transfer member relative to the working surface. Furthermore, the workstation may comprise a plurality of magnetic field generating elements, each magnetic field generating element being movable relative to the work surface between a first position remote from the at least one container and a second position adjacent to the at least one container.

Yet another aspect of the present disclosure provides a purification method, which may include: (a) providing a workstation having a working surface, a robotic manipulator movable relative to the working surface and carrying a fluid transfer member, and a plurality of magnetic field generating elements laterally spaced from one another to define a plurality of rows; (b) adding at least one culture medium having a target substance suspended therein to a plurality of containers or vessels; (c) adding a plurality of magnetic beads to the plurality of containers, the target substance temporarily binding to the plurality of magnetic beads; and (d) arranging the plurality of vessels in a plurality of rows defined between the plurality of magnetic field generating elements such that the plurality of magnetic field generating elements magnetically attract and hold the plurality of magnetic beads against an inner surface of a respective vessel of the plurality of vessels.

Drawings

It is believed that the present disclosure will be more fully understood from the following description in conjunction with the accompanying drawings. Some of the drawings may be simplified by omitting selected elements for the sake of more clearly showing other elements. The omission of such elements in some figures does not necessarily indicate the presence or absence of a particular element in any embodiment unless specifically indicated in the corresponding written description. Moreover, all of the accompanying drawings are not necessarily drawn to scale.

Fig. 1 is a top perspective view of an embodiment of a magnetic separator according to the principles of the present disclosure.

Fig. 2 is a front perspective view of the magnetic separator shown in fig. 1.

Fig. 3 is a rear perspective view of the magnetic separator shown in fig. 1.

FIG. 4 is a cross-sectional view of the magnetic separator illustrated in FIG. 1 with a container containing a fluid medium and magnetic beads.

Fig. 5 is a perspective side view of another embodiment of a magnetic separator according to the principles of the present disclosure.

Fig. 6A-6G schematically illustrate, in chronological order, the steps of one embodiment of a purification method according to the principles of the present disclosure.

FIG. 7 is a front plan view of a schematic representation of an embodiment of a workstation for magnetic separation according to the principles of the present disclosure.

Fig. 8 is a side plan view of a schematic representation of the workstation shown in fig. 7.

FIG. 9 is a front perspective view of an embodiment of a workstation according to the principles of the present disclosure.

Detailed Description

The present disclosure relates generally to devices and methods for separating a target substance, such as a protein, from a fluid medium in which the target substance is suspended. In a broad sense, the presently disclosed devices and methods include positioning one or more magnetic field generating elements adjacent to the exterior of one or more vessels containing a medium having a substance of interest. The plurality of magnetic beads may be submerged in the medium and may bind to the target substance. The one or more magnetic field generating elements may magnetically attract the magnetic beads bound to the target substance and statically fix or hold the magnetic beads against the inner surface of the one or more containers. The medium can then be removed from the one or more vessels, leaving the magnetic beads bound to the target substance. Subsequently, the magnetic beads may be subjected to a washing and/or elution procedure to release and/or extract the target substance from the magnetic beads. By moving only the one or more magnetic field generating elements away from the one or more containers, the magnetic beads can eventually be separated from the magnetic field and vice versa.

So configured, the devices and basic methods of the present disclosure advantageously simplify the process of isolating a substance of interest from a culture medium, and in certain embodiments, may eliminate or reduce the number of tasks that must be performed manually by a laboratory technician or other user. In addition, the presently disclosed apparatus and methods may provide the ability to process relatively large sample volumes without significantly increasing the footprint of the workstation required for the equipment associated with purification. Furthermore, the purification devices and methods of the present disclosure can reduce the likelihood of cross-contamination that may occur if one or more magnetic field generating elements are submerged in the sample medium.

Each of the foregoing components and associated methods will now be described in greater detail.

The devices and methods of the present disclosure can be used to separate a wide variety of target substances (e.g., molecules, molecular complexes, biomolecules, biomolecule complexes, proteins, protein complexes, peptides, nucleic acid ligands, pathogenic microorganisms, cells, etc.) from a wide variety of sample mixtures (e.g., cell cultures, blood, salvia, mucus, sweat, urine, stool, soil, food, etc.). Furthermore, a wide variety of magnetic beads may be used depending on the nature of the target substance to be sequestered and/or the medium in which the target substance is suspended. In some embodiments, the magnetic beads may have a spherical shape and have a silica-based paramagnetic core coated with a material that binds or conjugates with the target substance. The binding between the magnetic beads and the target substance may be achieved covalently, non-covalently, electrostatically, by hydrogen bonding, by van der waals forces, and/or by any other suitable molecular binding process. In at least one embodiment, the medium can be a cell culture or other biological mixture, the target substance can be an antibody or other protein, and the magnetic beads can be protein a magnetic beads. In another example, the medium may be a cell culture or other biological mixture, the target substance may be a polyhistidine-tagged protein, and the magnetic beads may be coated with nickel, zinc, copper, or cobalt.

A non-limiting list of examples of types of magnetic beads that can be implemented in the presently disclosed purification systems and methods includes: affinity magnetic beads (e.g., amine magnetic beads, aldehyde magnetic beads, carboxyl magnetic beads, CDI magnetic beads, DVS magnetic beads, DADPA magnetic beads, epoxy magnetic beads, hydrazide magnetic beads, hydroxyl magnetic beads, iodoacetyl magnetic beads, NHS magnetic beads, thiol magnetic beads, tosyl magnetic beads, thiol magnetic beads, silica magnetic beads, IDA magnetic beads, etc.); reversed-phase magnetic beads (e.g., C4 magnetic beads, C8 magnetic beads, C18 magnetic beads, cyanopropyl magnetic beads, phenyl magnetic beads, diphenyl magnetic beads, etc.); ion exchange type magnetic beads (e.g., DEAE magnetic beads, PSA magnetic beads, SAX magnetic beads, WCX magnetic beads, SCX magnetic beads, hydroxyapatite magnetic beads, etc.); antibody-purifying type magnetic beads (e.g., protein a magnetic beads, protein G magnetic beads, protein a/G magnetic beads, protein L magnetic beads, fast IgG pure magnetic beads, antigenic peptide magnetic beads, fast IgM pure magnetic beads, anti-IgG magnetic beads, fast IgA pure magnetic beads, thiophilic magnetic beads, etc.); antibody-immobilized magnetic beads (e.g., protein a magnetic beads, protein G magnetic beads, protein a/G magnetic beads, protein L magnetic beads, epoxy-activated magnetic beads, aldehyde-terminated magnetic beads, hydrazide-terminated magnetic beads, carboxyl-terminated magnetic beads, iodoacetyl-activated magnetic beads, thiol-activated magnetic beads, etc.); recombinant protein purification type magnetic beads (e.g., Ni + charged magnetic beads, Co + charged magnetic beads, maltose magnetic beads, calmodulin magnetic beads, etc.); peptide-immobilized magnetic beads (e.g., epoxy-activated magnetic beads, aldehyde-terminated magnetic beads, carboxyl-terminated magnetic beads, amine-terminated magnetic beads, iodoacetyl-activated magnetic beads, thiol-activated magnetic beads, etc.); magnetic beads for DNA or RNA purification; magnetic beads for removing endotoxin; magnetic beads for removing a large amount of proteins; and/or EDTA magnetic beads.

The term "magnetic" as used herein is defined to encompass any element that is magnetic, paramagnetic, and/or ferromagnetic. Thus, the magnetic beads may be magnetic beads, paramagnetic beads, ferromagnetic beads, or any combination thereof.

In some embodiments, the density of the magnetic beads can be greater than the density of the culture medium such that when the magnetic beads are submerged in the sample mixture, the magnetic beads sink to the bottom of the container holding the culture medium. In other embodiments, the density of the magnetic beads can be less than or equal to the media such that the magnetic beads float or partially float in the media.

Fig. 1-3 illustrate one embodiment of a magnetic separator 10 according to the principles of the present disclosure. The magnetic separator 10 generally includes a frame 12 for mounting magnetic field generating elements 14 and 16 at a distance X1 from each other. An opening 18 may be formed in the frame 12 and sized so that one or more receptacles 20 may be inserted through the opening 18 into the space between the magnetic field generating elements 14 and 16, as shown in fig. 3. In certain embodiments, the container 20 may be attached to or held by a non-magnetic support bracket 50 (see fig. 4) that may also be positioned between the mounting magnetic field generating elements 14 and 16.

In general, the frame 12 serves to maintain the spaced distance X1 between the magnetic field generating elements 14 and 16 despite the presence of magnetic attraction and/or repulsion forces between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the receptacle 20. Securing the magnetic field generating elements 14 and 16 at the set distance X1 by the frame 12 may not require a user to set or otherwise handle the magnetic field generating elements 14 and 16, which may be cumbersome and may be unsafe in situations where the magnetic tension between the magnetic field generating elements 14 and 16 is relatively strong. Moreover, by not having to make direct contact with the magnetic field generating elements 14 and 16, there is less risk of damage to the memory or other sensitive components of the instrument or personal electronic device carried by the user.

In some embodiments, the lateral distance X1 separating the first and second magnetic field generating elements 14 and 16 may be in a range of between about (e.g., ± 10%) 3 and 36 inches, or in a range of between about (e.g., ± 10%) 3 and 24 inches, or in a range of between about (e.g., ± 10%) 3 and 18 inches, or in a range of between about (e.g., ± 10%) 3 and 12 inches, or greater than or equal to about (e.g., ± 10%) 1 inch, or greater than or equal to about (e.g., ± 10%) 2 inches, or greater than or equal to about (e.g., ± 10%) 3 inches, or greater than or equal to about (e.g., ± 10%) 4 inches, or equal to about (e.g., ± 10%) 3.4 inches.

In the present embodiment, the separation distance X1 between the magnetic field generating elements 14 and 16 is not adjustable. However, in alternative embodiments, the frame 12 may have an adjustable width, allowing the user to adjust the distance X1 between the magnetic field generating elements 14 and 16, and then lock the magnetic field generating elements 14 and 16 in place.

Referring to fig. 1-4, the frame 12 may be formed from a first vertical sidewall 30, a second vertical sidewall 32, and a horizontal bottom wall 34. The first and second vertical sidewalls 30, 32 may be arranged parallel to each other as shown, or in an alternative embodiment, may be arranged at a non-parallel angle with respect to each other. The horizontal bottom wall 34 may extend between the first and second vertical sidewalls 30 and 32 and may provide the necessary structural support to prevent the first and second vertical sidewalls 30 and 32 from moving relative to each other due to the magnetic attraction and/or repulsion forces present between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the container 20. As shown in fig. 1-3, additional structural support may be provided by one or more beams 34a-d extending between the first and second vertical sidewalls 30, 32. In an alternative embodiment, the bottom wall 34 may be omitted and only one or more beams 34a-d may be included to keep the first and second vertical sidewalls 30, 32 apart. In a further alternative embodiment, the beams 34a-d may be omitted, and only the bottom wall 34 may be included to maintain the separation distance of the first and second vertical sidewalls 30, 32.

As depicted in fig. 1-3, the frame 12 does not include a front wall, a top wall, or a rear wall. Instead of a front wall, an opening 18 (i.e., a front opening) is defined between the first and second vertical sidewalls 30, 32. Similarly, instead of a top wall and a rear wall, a top opening 26 and a rear opening 28 are defined between the first and second vertical sidewalls 30 and 32, respectively. In alternative embodiments, a top wall and/or a rear wall may be included such that the openings are only the front and rear openings 18, 28, or only the front and top openings 18, 26, or only the front opening 18. In a further alternative embodiment, the top opening 26 may be the only opening formed in the frame 12. In the illustrated embodiment, the front opening 18 is separated from the top opening 26 by a beam 34a, and the top opening 26 is separated from the rear opening 28 by a beam 34 b. In other embodiments, the front opening 18, the top opening 26, and the rear opening 28 may be continuous with one another (see, e.g., fig. 5) with the beams 34a and 34b omitted.

Moreover, while the vertical side walls 30 and 32 and the horizontal bottom wall 34 are illustrated as solid structures extending continuously along the entire length of the separator 10, in alternative embodiments, one or more of the vertical walls 30 and 32 and/or the horizontal bottom wall 34 may be formed from one or more struts or trusses with gaps therebetween, or with one or more cut-out sections, to reduce the weight of the frame 12.

The frame 12 may be constructed of a rigid material including, but not limited to, some type of metal and/or plastic. The rigidity of the frame 12 should be such that the magnetic attraction and/or repulsion forces present between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the receptacle 20 do not cause the frame 12 to deform. In some embodiments, the frame 12 may be constructed of, for example, a non-magnetic material such as plastic and/or aluminum; however, in other embodiments, the frame 12 may be made of a magnetic material such as ferritic stainless steel.

With continued reference to fig. 1-4, the first magnetic field generating element 14 may be rigidly secured to an inward facing surface 36 of the first vertical sidewall 30, and the second magnetic field generating element 16 may be rigidly secured to an inward facing surface 38 of the second vertical sidewall 32. In some embodiments, the first magnetic field generating element 14 and/or the second magnetic field generating element 16 may be rigidly fixed, for example, with fasteners such as bolts and/or screws. Additionally, in some embodiments, each of the first and/or second magnetic field generating elements 14 and 16 can be housed in a respective cage or other housing 44 or 46 that is rigidly secured to the inwardly facing surface 36 of the first vertical side wall 30 or the inwardly facing surface 38 of the second vertical side wall 32. Each cage 44 and 46 can have side openings 48 or 50 to allow the first or second magnetic field generating element 14 or 16 to be slidably inserted into or removed from its respective cage 44 or 46.

Each of the magnetic-field-generating elements 14 and 16 may be mounted such that its downwardly-facing or bottom surface is spaced in the vertical direction from the horizontal bottom wall 34, or if the horizontal bottom wall 34 is omitted, from the surface on which the frame 12 rests, by a distance X2. As discussed below, this may provide space or clearance for the sides of the non-magnetic support frame to be inserted under the first and second magnetic field generating elements 14 and 16, respectively.

As shown in fig. 4, the first and second magnetic field generating elements 14, 16 project inwardly from inwardly facing surfaces 36, 38 of the first and second vertical sidewalls 30, 32, respectively. In alternative embodiments, the first and second magnetic field generating elements 14, 16 may be positioned in respective grooves or recesses formed in the inwardly facing surfaces 36, 38 of the first and second vertical sidewalls 30, 32, respectively. In this way, the first and second magnetic field generating elements 14 and 16 may be flush with the inwardly facing surfaces 36 and 38, respectively.

In some embodiments, as shown in fig. 4, the first and second magnetic field generating elements 14, 16 may have a generally flat or planar shape with a rectangular cross-section. However, other shapes and cross-sections are possible. Also, magnetic field generating elements other than those depicted in the figures may also be included. Additionally, in some embodiments, only a single magnetic field generating element may be included, such that one of the vertical sidewalls 30 or 32 is free of magnetic field generating elements.

In some embodiments, each of the magnetic field generating elements 14 and 16 may be comprised of a respective permanent magnet configured to generate its own permanent magnetic field. The maximum magnetic pull force of each permanent magnet may be in a range between about (e.g., ± 10%) 50 to 1000 newtons (N), or about (e.g., ± 10%) 100 to 800N, or about (e.g., ± 10%) 100 to 700N, or about (e.g., ± 10%) 150 to 600N, or about (e.g., ± 10%) 200 to 500N, or about (e.g., ± 10%) 200 to 450N, or greater than or equal to about (e.g., ± 10%) 50N, or greater than or equal to about (e.g., ± 10%) 100N, or greater than or equal to about (e.g., ± 10%) 150N, or greater than or equal to about (e.g., ± 10%) 200N, or greater than or equal to about (e.g., ± 10%) 250N. In some embodiments, the total combined magnetic pull force of the permanent magnets may be greater than or equal to about (e.g., ± 10%) 500N, or greater than or equal to about (e.g., ± 10%) 1000N, or greater than or equal to about (e.g., ± 10%) 1500N, or greater than or equal to about (e.g., ± 10%) 2000N, or greater than or equal to about (e.g., ± 10%) 2500N. In some embodiments, the permanent magnets that make up the magnetic field generating elements 14 and 16 may be nickel plated neodymium block magnets. In an alternative embodiment, each of the magnetic field generating elements 14 and 16 may be constituted by a respective electromagnet configured to generate a magnetic field when supplied with an electric current.

Referring to fig. 4, the magnetic separator 10 may include a non-magnetic support 50 that is separate from and movable relative to the frame 12. In general, the non-magnetic support 50 may be configured to hold a plurality of containers 20 containing a culture medium 54 in which the target substance T is initially suspended. In some embodiments, the non-magnetic support 50 may be configured to prevent lateral movement of the containers 20 relative to each other due to magnetic attraction and/or repulsion forces present between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the containers 20. This aspect of the non-magnetic support 50 may be achieved by various configurations, including, for example: forming a non-magnetic support 50 having a plurality of apertures, each aperture sized to receive one of the containers 20; or by rigidly securing the container 20 to the non-magnetic support 50 by one or more fasteners or adhesives; alternatively, as shown in the embodiment illustrated in FIG. 5, the container 120 is integrally joined with the support frame 150 such that they form a single unitary structure. The non-magnetic support 50 may be constructed of any non-magnetic material including, for example, plastic or glass, such that it is not magnetically attracted or repelled by the first and second magnetic field generating elements 14 and 16. The container 20 may also be constructed of a non-magnetic material including, for example, glass or plastic.

In some embodiments, the non-magnetic support 50 may be configured to hold the containers 20 in one, two, three, or more rows. Additionally, in some embodiments, each row of containers 20 formed from non-magnetic support shelves 50 may extend longitudinally in a direction parallel to the first and second vertical sidewalls 30, 32.

The non-magnetic support 50 may have a width or otherwise be sized such that it may be inserted in a generally horizontal direction through the opening 18 in the front side of the frame 12. Such insertion may cause the container 20 to be positioned in the space between the first and second magnetic field generating elements 14, 16. In some embodiments, such as the embodiment shown in fig. 4, the width W1 of the non-magnetic support 50 may be greater than the distance X1 separating the first and second magnetic field generating elements 14 and 16. In such embodiments, the height H1 of the non-magnetic support 50 may be less than the vertical distance X2 separating the bottom surfaces of the first and second magnetic field generating elements 14, 16 from the horizontal bottom wall 34 of the frame 12. Thus, the first and second lateral ends 56, 58 of the non-magnetic support 50 may fit under the first and second magnetic field generating elements 14, 16, respectively. This configuration may advantageously limit lateral movement of the non-magnetic support 50 caused by the inertia of the magnetic field generating elements 14 and 16 magnetically pulling against the magnetic beads 40 on the inner surface of their respective receptacle 20. This is because the first and second lateral ends 56 and 58 of the non-magnetic support bracket 50 may abut the inwardly facing surfaces 36 and 38 of the first and second vertical sidewalls 30 and 32, respectively, thereby preventing significant lateral movement of the non-magnetic support bracket 50.

Turning to fig. 5, another embodiment of a magnetic separator 110 in accordance with the principles of the present disclosure is illustrated. Magnetic separator 110 is similar to magnetic separator 10 except that it does not include a beam extending between vertical walls 130 and 132 of frame 112. Elements of the magnetic separator 110 illustrated in fig. 5 that are the same as or similar to elements of the magnetic separator 10 illustrated in fig. 1-4 are indicated by the same reference numerals increased by 100. The description of these similar components is omitted for the sake of brevity.

The absence of beams extending between the vertical walls 130 and 132 of the frame 112 allows the front opening 118, the top opening 126, and the rear opening 128 to be continuous with one another. Accordingly, the space between the first and second magnetic field generating elements 114 and 116 may be more easily accessible to a user, and there may be more than one insertion path for the non-magnetic support stand 150.

A method of using the magnetic separator 10 in a purification process will now be described with reference to fig. 6A to 6G. Fig. 6A to 6G are schematic side views, and various elements of the magnetic separator 10 are omitted. The omission of these elements is not to be understood as meaning that they must be lost from the magnetic separator 10. Also, the magnetic separator 110 may be used in a process similar to that described in fig. 6A-6G.

As an initial step, each vessel 20 or individual vessel 20 may be filled with a volume of medium 54 comprising the target substance T. Next, as shown in fig. 6A, magnetic beads 40 can be added to the container 20 and allowed to interact with the culture medium 54 for a period of time (e.g., minutes, an hour, hours, a day, days, etc.). During this incubation, the magnetic beads 40 may bind to the target substance T, thereby separating the target substance T from the rest of the culture medium 54 (see fig. 6B). As previously mentioned, the binding between the magnetic beads 40 and the target substance T may be achieved covalently, non-covalently, electrostatically, by hydrogen bonding, van der waals forces, and/or any other suitable molecular binding process. In some embodiments, the medium 54 may be stirred or agitated during incubation to promote binding between the target species T and the magnetic beads 40. Additionally, in some embodiments, the container 20 may be placed in the magnetic separator 10 during the period of time that the magnetic beads 40 are bound to the target substance T.

Next, if not completed, the container 20 may be secured to the non-magnetic support stand 50, and the non-magnetic support stand 50 may be inserted between the first and second vertical sidewalls 30, 32 of the frame 12 such that the container 20 is positioned between the first and second magnetic field generating elements 14, 16, as shown in fig. 6C. This step may require the user to manually move the non-magnetic support 50 with the container 20 disposed thereon from a first position external to the frame 12 to a second position between the first and second vertical sidewalls 30, 32 of the frame 12. In some embodiments, this movement may include inserting the non-magnetic support 50 and the receptacle 20 through the front opening 18 of the frame 12 in a horizontal direction. In some embodiments, the first and second lateral ends 56 and 58 of the non-magnetic support 50 may slide or otherwise fit under the first and second magnetic field generating elements 14 and 16 during insertion. In alternative embodiments, the non-magnetic support 50 and the receptacle 20 may be inserted through the top opening 26 in a vertically downward direction or through the rear opening 28 in a horizontal direction.

As shown in fig. 6C, when the container 20 is positioned between the first vertical sidewall 30 and the second vertical sidewall 32, the proximity of the first magnetic field generating element 14 and the second magnetic field generating element 16 may allow them to magnetically attract and hold the magnetic beads 40 against the inner surface 60 of their respective sidewall 61 of the container 20. The friction between the magnetic bead 40 and the inner surface 60 of the container 20 may effectively immobilize or immobilize the magnetic bead 40 relative to the container 20 such that the magnetic bead 40 is prevented or inhibited from moving during subsequent fluid removal and addition steps.

Next, as shown in fig. 6D, the purification method may include aspirating or removing the culture medium 54 from the container 20 via a fluid transfer member 62, such as a pipette or other fluid conduit. The distal end or mouth of fluid transfer member 62 may be inserted through a top opening in frame 12 and then into an opening 64 formed in the top of one of containers 20, thereby submerging it in media 54. The mouth of fluid transfer member 62 may be positioned adjacent to or otherwise in close proximity to bottom wall 66 of container 20 such that fluid transfer member 62 may aspirate all or substantially all of media 54 in container 20. The magnetic beads 40 are not removed in this step, since they are held against the inner surface 60 of the side wall 61 of the container 20 by the magnetic pulling force of the first and/or second magnetic field generating element 14, 16. In addition, the target substance T bound to the magnetic beads 40 also remains in the container 20. This step of removing the medium 54 with the target substance T removed may be repeated for all vessels 20, or in some embodiments, performed simultaneously for all vessels 20 with multiple fluid transfer members.

After removing the media 54, a wash fluid 70 (e.g., a saline solution) may be added to the container 20 via the fluid transfer member 62 or another fluid conduit to clean the interior surface 60 of the container 20 and/or any remaining magnetic beads 40 of the media 54, as shown in fig. 6E. However, the washing fluid 70 is not able to remove the target substance T from the magnetic beads 40. The volume of the wash fluid 70 added to the containers 20 may be equal to or less than the maximum volume of the respective container 20 (e.g., a fraction thereof). Alternatively, just prior to or just after adding the washing fluid 70 to the receptacle 20, the receptacle 20 or the receptacle 20 of the entire rack may be removed from the frame 12 such that the receptacle 20 is not positioned between the first and second magnetic field generating elements 14, 16. In the absence of the relatively strong magnetic field of the magnetic field generating elements 14 and 16, the magnetic beads 40 may be freely dispersed in the washing fluid 60 and may be suspended in the washing fluid 60. Allowing the magnetic beads 40 to disperse in this manner may increase the efficiency of the washing process and/or allow stirring or other agitation of the magnetic beads 40 in the washing fluid 70. After washing is complete, the washing fluid 70 may be removed from the container 20 via the fluid transfer member 62 or other fluid conduit.

In alternative embodiments, one or more or all of the steps associated with the above-described wash fluid may be omitted.

Next, as shown in fig. 6F, an eluent 72 (e.g., a liquid elution solution) may be added to the container 20 through the fluid transfer component 62 or another fluid conduit to release the target species T from the magnetic beads 40. If not, the receptacle 20, or the receptacle 20 of the entire rack is removed from the frame 12 just prior to or just after adding the eluant 72 to the receptacle 20 such that the receptacle 20 is not positioned between the first and second magnetic field generating elements 14, 16. In the absence of the relatively strong magnetic fields of magnetic field generation 14 and 16, the magnetic beads 40 may be freely dispersed in the eluant 72 and may be suspended in the eluant 72. Allowing the magnetic beads 40 to disperse in this manner may increase the efficiency of the elution process and/or allow stirring or other agitation of the magnetic beads 40 in the eluate 72. Alternatively, this elution step may be performed while the container 20 is positioned between the first and second magnetic field generating elements 14, 16.

After a period of incubation, the eluate 72 and the target substance T bound to the eluate 72 can be removed from the container 20 via the fluid transfer member 62 or another conduit, and subsequently discharged into an external container. If not, the container 20 may be repositioned between the first and second magnetic field generating elements 14, 16 to immobilize the magnetic beads 40 against the inner surface 60 of their respective container 20, just prior to this fluid removal step. The above steps can then be repeated for another vessel or batch of vessels containing another medium or mixture that needs to be purified.

It is noted that any of the aforementioned steps of positioning the container 20 between the first and second magnetic field generating elements 14, 16 and/or removing the container 20 from between the first and second magnetic field generating elements 14, 16 may be performed manually by a user, such as a laboratory technician. Also, the user may manually perform any fluid addition or removal steps by means of, for example, a pipette. Additionally, it should be noted that magnetic separator 10 is sized, otherwise configured, such that it can be supported on a variety of standard laboratory tables and/or table tops.

Turning to fig. 7 and 8, a schematic representation of a workstation 200 for separating a target substance from a medium in which the target substance is suspended is illustrated, in accordance with the principles of the present disclosure. The workstation 200 is similar in some respects to the magnetic separator embodiments described above in that one or more magnetic field generating elements are used to immobilize magnetic beads that are immersed in the culture medium and bind to the target substance during fluid removal steps, such as during removal of the culture medium, removal of the wash fluid, and/or removal of the elution fluid. However, the workstation 200 differs from the previous embodiments of the magnetic separator in that one or more magnetic field generating elements are moved between different positions during the washing purification process while the container remains stationary. Further, unlike the magnetic separator described above, the workstation 200 comprises a robotic manipulator configured to move at least one fluid transfer member relative to the container for removing and/or adding fluid.

In general, workstation 200 is sized or otherwise configured such that it may be supported on a variety of standard laboratory tables and/or tables. Referring to fig. 7-9, workstation 200 may generally include a frame 210 including a horizontally disposed and upwardly facing work surface 212, a fluid transfer member 214, a robotic manipulator 216, and one or more magnetic field generating elements 218 a-e. Optionally, the workstation 200 may include one or more linear actuators 220a-e connected to one or more magnetic field generating elements 218a-e, one or more pumps 222a-b in fluid communication with the fluid transfer member 214, a multi-position valve 224 in fluid communication with one or more pumps 222a-b, a waste container or drain 226, one or more auxiliary containers 228a-c containing one or more eluents, washing fluids, and/or other fluids, and/or a control unit 230.

As shown in fig. 7, the magnetic field generating elements 218a-e may be laterally spaced apart from one another at regular intervals in a horizontal direction to define a plurality of rows 232a-d for slidably receiving a plurality of receptacles 240. In some embodiments, the lateral distance X3 separating adjacent ones of the magnetic field generating elements 218a-e may be in a range between about (e.g., ± 10%) 3 and 10 inches, or may be in a range between about (e.g., ± 10%) 3 and 8 inches, or in a range between about (e.g., ± 10%) 3 and 6 inches, or in a range between about (e.g., ± 10%) 3 and 5 inches, or greater than or equal to about (e.g., ± 10%) 1 inch, or greater than or equal to about (e.g., ± 10%) 2 inches, or greater than or equal to about (e.g., ± 10%) 3 inches, or greater than or equal to about (e.g., ± 10%) 4 inches, or equal to about (e.g., ± 10%) 3.4 inches.

Each vessel 240 may be initially filled with a culture medium 242 in which the target substance T is suspended. Each container 240 may have an opening 244 formed at a top thereof, an inner surface 246, a sidewall 248, and a bottom wall 250 connected to the sidewall 248. The medium 242 and the target substance T may be similar to the medium and the target substance discussed above. Within each row 232a-d, the containers 240 may be held by a non-magnetic support 252, which in some embodiments may be configured in a similar manner to the non-magnetic support discussed above.

A plurality of magnetic beads 260 may be submerged in the media 242 in each vessel 240. The magnetic beads 260 may be constructed in a similar manner to the magnetic beads 40 described above, and may be configured to bind to the target substance T suspended in the medium 242.

Each magnetic field generating element 218a-e may extend longitudinally along a longitudinal axis a (see fig. 8). Additionally, in some embodiments, the longitudinal axes A of the magnetic field generating elements 218a-e may be parallel to one another. The length L2 of each magnetic field generating element 218a-e may be parallel to the longitudinal axis a of the respective magnetic field generating element, and the width W2 of the magnetic field generating element 218a-e may be perpendicular to the longitudinal axis a of the respective magnetic field generating element. In some embodiments, length L2 may be in the range of about (e.g., ± 10%) 10 to 48 inches, or in the range of about (e.g., ± 10%) 10 to 36 inches, or in the range of about (e.g., ± 10%) 10 to 30 inches, in the range of about (e.g., ± 10%) 10 to 24 inches, or equal to or greater than about (e.g., ± 10%) 10 inches, or equal to or greater than about (e.g., ± 10%) 20 inches, or equal to or greater than about (e.g., ± 10%) 30 inches. In some embodiments, the width W2 may be in the range of about (e.g., ± 10%) 0.5 to 6 inches, or in the range of about (e.g., ± 10%) 0.5 to 4 inches, or in the range of about (e.g., ± 10%) 0.5 to 3 inches, or in the range of about (e.g., ± 10%) 0.5 to 2 inches, or equal to or greater than about (e.g., ± 10%) 0.5 inches, or equal to or greater than about (e.g., ± 10%) 1 inch, or equal to or greater than about (e.g., ± 10%) 1.5 inches, or equal to or greater than about (e.g., ± 10%) 2 inches, or equal to or greater than about (e.g., ± 10%) 2.5 inches.

Each magnetic field generating element 218a-e is movable back and forth relative to the working surface 212 between a first, or non-operative, position away from the respective row of containers 240 and a second, or operative, position adjacent (e.g., immediately adjacent) to the respective row of containers 240. Fig. 8 schematically illustrates in solid lines one of the magnetic field generating elements 218a-e occupying its operative position. Fig. 8 also illustrates in dashed lines the non-operative position of this one of the magnetic field generating elements 218 a-e. In the present embodiment, the workstation 200 includes a plurality of linear actuators 220a-e, each configured to independently reciprocate a respective one of the magnetic field generating elements 218a-e in a linear horizontal direction between its operative and inoperative positions. In other embodiments, a single linear actuator may be used to simultaneously move all of the magnetic field generating elements 218a-e back and forth between their operative and inoperative positions in common. Each linear actuator 220a-e may include at least one of a hydraulic cylinder, a pneumatic cylinder, or an electric motor. In embodiments that include a hydraulic or pneumatic cylinder, a reciprocating piston may connect the hydraulic or pneumatic cylinder to a respective one of the magnetic field generating elements 218 a-e. In embodiments that include a rotary electric motor, a rack and pinion arrangement, pulley, or gear system may be used to convert the rotary motion output generated by the electric motor into linear motion of a respective one of the magnetic field generating elements 218 a-e.

In the embodiment shown in FIG. 8, each linear actuator 220a-e takes the form of a pneumatic linear slider. As such, each linear actuator 220a-e may include a pneumatic cylinder 221a-e for receiving pressurized gas, and a piston or carriage 223a-e that translates back and forth by pressurizing different ends of the pneumatic cylinder 221 a-e. As shown in FIG. 8, each pneumatic cylinder 221a-e may be disposed within the interior space of the frame 210 vertically below the working surface 212. Each of the load bearing elements 223a-e may be mechanically coupled to a respective one of the magnetic field generating elements 218a-e through a respective opening 225a-e formed in the working surface 212. Although fig. 8 illustrates only the linear actuator 220e and its connection to the magnetic field generating element 218e, the other magnetic field elements 218a-d and linear actuators 220a-d may be configured in a similar manner and are not illustrated for purposes of brevity. In an alternative embodiment, the linear actuator 220e may be configured to move all of the magnetic field generating elements 218a-e together.

Although the magnetic field generating elements 218a-e of the present embodiment may move in a horizontal direction parallel to the working surface 212, in alternative embodiments, each magnetic field generating element 218a-e may move in a vertical direction that is perpendicular or otherwise non-parallel to the working surface 212. In such alternative embodiments, a plurality of openings may be formed in the working surface 212, and each magnetic field generating element 218a-e may be configured to reciprocally retract into and extend out of a respective one of the openings. Here, the retracted position (which may be below the working surface 212) may correspond to an inoperative position, and the extended position (which may be above the working surface 212) may correspond to an operative position.

In some embodiments, such as illustrated in FIG. 9, the linear actuators 220a-e may be omitted. In such embodiments, movement of the magnetic field generating elements 218a-e may be accomplished by a user manually moving each magnetic field generating element 218a-e in a horizontal and/or vertical direction between its operative and inoperative positions.

When the magnetic field generating elements 218a-e occupy their respective operating positions, they can magnetically attract and hold the magnetic beads 260 against the inner surface 246 of the sidewall 248 of the container 240. The friction between the magnetic beads 260 and the inner surface 246 of the container 240 may effectively immobilize or immobilize the magnetic beads 260 relative to their respective container 240 such that the magnetic beads 260 are prevented or prevented from moving during subsequent fluid removal and/or addition steps.

In some embodiments, each magnetic field generating element 218a-e may be comprised of a respective permanent magnet configured to generate its own permanent magnetic field. The maximum magnetic pull force of each permanent magnet may be in a range between about (e.g., ± 10%) 50 and 1000 newtons (N), or in a range between about (e.g., ± 10%) 100 and 800N, or in a range between about (e.g., ± 10%) 100 and 700N, or in a range between about (e.g., ± 10%) 150 and 600N, or in a range between about (e.g., ± 10%) 200 and 500N, or in a range between about (e.g., ± 10%) 200 and 450N, or in a range between about (e.g., ± 10%) 250 and 350N, or greater than or equal to about (e.g., ± 10%) 50N, or greater than or equal to about (e.g., ± 10%) 100N, or greater than or equal to about (e.g., ± 10%) 150N, or greater than or equal to about (e.g., 10%) 200N, or greater than or equal to about (e.g., ± 10%) 250N, or equal to about (e.g., ± 10%) 289N. In some embodiments, the total combined magnetic pull force of the permanent magnets may be greater than or equal to about (e.g., ± 10%) 500N, or greater than or equal to about (e.g., ± 10%) 1000N, or greater than or equal to about (e.g., ± 10%) 1500N, or greater than or equal to about (e.g., ± 10%) 2000N, or greater than or equal to about (e.g., ± 10%) 2500N, greater than or equal to about (e.g., ± 10%) 5000N, or greater than or equal to about (e.g., ± 10%) 7000N, or greater than or equal to about (e.g., ± 10%) 7500N, or equal to about (e.g., ± 10%) 7225N. In some embodiments, the permanent magnets comprising the magnetic field generating elements 218a-e may be nickel plated neodymium block magnets. In an alternative embodiment, each magnetic field generating element 218a-e may be comprised of a respective electromagnet configured to generate a magnetic field when supplied with an electrical current.

With continued reference to fig. 7-9, robotic manipulator 216 may be configured to automatically move fluid transfer member 214 between different positions relative to working surface 212, which may or may not be predefined or preprogrammed. The robotic manipulator 216 may include one or more electric motors controlled by the control unit 230. In the illustrated embodiment, the robotic manipulator 216 takes the form of a cartesian robot that can move independently in each of the x, y, and z directions. In the present embodiment, the x-direction and the y-direction are horizontal linear directions, and the z-direction is a vertical linear direction. The robotic manipulator 216 may include separate electric motors and separate track members for achieving each direction of movement. In other embodiments, the robotic manipulator 216 may be capable of movement in only two directions (e.g., only the x-direction and the y-direction, or only the x-direction and the z-direction, or only the y-direction and the z-direction) or only in a single linear direction. In yet further embodiments, the robotic manipulator 216 may take the form of a robotic arm capable of compound bending, linear, and/or rotational motion.

Still referring to fig. 7-9, the robotic manipulator 216 may include a mounting plate 262 for mounting the fluid transfer member 214, the pumps 222a-b, the multi-position valve 224, and/or other components to be carried by the robotic manipulator 216. The fluid transfer member 214 may be mechanically connected (e.g., fastened with fasteners) to the mounting plate 262 and may extend downward in a vertical direction from the mounting plate 262. In the present embodiment, the fluid transfer member 214 includes two parallel vertical fluid conduits 264a and 264b (e.g., flexible or rigid plastic tubes) that are spaced apart from each other by a horizontal distance. The fluid conduit 264a may be in fluid communication with the pump 222a, and the fluid conduit 264b may be in fluid communication with the pump 222 b. The two fluid conduits 264a and 264b may be separated by a horizontal distance equal to the horizontal distance separating the centers of any two adjacent rows 232 a-d. Additionally, in some embodiments, the horizontal distance separating the two fluid conduits 264a and 264b may be adjustable to account for different sized containers 240. In alternative embodiments, fluid transfer member 214 may include only a single vertical fluid conduit, or three or more vertical fluid conduits.

In general, each pump 222a-b may be configured to remove and/or add fluid to a respective container in 240 via fluid transfer member 214. Each pump 222a-b may be driven by any suitable means, including but not limited to an electric motor and/or a source of pressurized hydraulic fluid and/or gas. Each pump 222a-b may operate at a variable speed or a single speed depending on the specifications of the purification process. In some embodiments, the operation of one or more pumps 222a-b may be electronically controlled by control unit 230 according to, for example, programmable instructions stored in a memory of control unit 230. Alternatively or additionally, each pump 222a-b may be operated by a user (e.g., a laboratory technician) manually actuating an ON/OFF switch and/or a rotational speed knob. In some embodiments, such as on the one depicted in fig. 9, one or more pumps 222a-b may be configured as positive displacement pumps (such as peristaltic pumps) and capable of pumping fluids containing suspended solids (e.g., magnetic beads 260) without causing damage to these solids. In an alternative embodiment, one or more of the pumps 222a-b may be centrifugal pumps (such as radial pumps) that employ a rotating impeller to create a vacuum to move the fluid. Additionally, in some embodiments, each pump 222a-b may be reversible.

The multi-position valve 224 may be configured to selectively fluidly connect the pump 222a and/or the pump 222b to one or more of the auxiliary containers 228a-c, the drain 226, and/or other elements. Depending on the number of auxiliary vessels, vents, etc., the multi-position valve 224 may be a 3-way valve, a 4-way valve, a 5-way valve, a 6-way valve, a 7-way valve, an 8-way valve, or any other valve having any number of selectively openable apertures. In some embodiments, the operation of the multi-position valve 224 may be electronically controlled by the control unit 230 according to, for example, programmable instructions stored in a memory of the control unit 230. The multi-position valve 224 may incorporate one or more solenoids for opening and closing the orifices of the multi-position valve 224 in response to command signals from the control unit 230.

In general, control unit 230 may be electrically connected to at least robotic manipulator 216, pumps 222a-b, and linear actuators 220a-e (if included), such that control unit 230 may send and/or receive electrical control signals to and/or from these components. The control unit 230 may include a processor (e.g., a microprocessor), a memory (e.g., Random Access Memory (RAM), non-volatile memory (such as a hard disk executable by the processor), flash memory, removable memory, non-removable memory, etc.) for storing tangible, non-transitory computer-readable instructions, a communication unit, a display, and an input device (e.g., a keyboard, a keypad, a touch screen, etc.). In some embodiments, the control unit 230 may be a programmable logic controller. The control unit 230 may be programmed to perform the purification process according to a predefined specification of the user. In some embodiments, the control unit 230 may perform steps of the purification process, such as activating the robotic manipulator 216 to move the fluid transfer member 214 and/or activating one or more linear actuators 220a-e to move one or more magnetic field generating elements 218a-e, in response to sensor data received from one or more sensors included in or used in conjunction with the workstation 200 and indicative of characteristics (e.g., volume, temperature, weight, pH, etc.) of the medium 242 and/or the target substance T, timers, analog or digital inputs from an operator, and/or any other relevant detectable event or occurrence.

The method of using the workstation 200 in the purification process will now be described. As an initial step, each vessel 240 or individual vessels 240 may be filled with a volume of culture medium 242 comprising the target substance T. Next, the magnetic beads 260 can be added to the container 240 and allowed to interact with the culture medium 242 for a period of time (e.g., minutes, an hour, hours, a day, days, etc.). During this incubation, the magnetic beads 260 can bind to the target substance T, thereby separating the target substance T from the rest of the medium 242 (similar to fig. 6B). As previously mentioned, the binding between the magnetic beads 260 and the target substance T may be achieved covalently, non-covalently, electrostatically, by hydrogen bonding, van der waals forces, and/or any other suitable molecular binding process. In some embodiments, the medium 242 may be stirred or agitated during incubation to promote binding between the target species T and the magnetic beads 242.

Next, if not completed, the containers 240 may be secured in groups to a plurality of non-magnetic support frames 252. Each loaded non-magnetic support bracket 252 may then be inserted horizontally between a corresponding pair of magnetic field generating elements 218a-e on top of the work surface 212. In doing so, each receptacle 240 may be disposed in one of the rows 232a-d defined between adjacent ones of the magnetic field generating elements 218 a-e. This step may include the user manually positioning each non-magnetic support 252 in a respective one of the rows 232 a-d. This insertion step may be performed with each of the magnetic field generating elements 218a-e positioned in their respective operating positions (see fig. 9), or alternatively with each of the magnetic field generating elements 218a-e positioned in their respective operating positions. In the latter case, the magnetic field generating elements 218a-e may be moved to their respective operating positions after the non-magnetic support 252 is placed on the work surface 212.

The proximity of the magnetic field generating elements 218a-e may allow them to magnetically attract and hold the magnetic beads 260 against the inner surface 246 of the sidewall 248 of their respective receptacle 240. The friction between the magnetic beads 260 and the inner surface 246 of the container 240 may effectively immobilize or immobilize the magnetic beads 260 relative to the container 240, such that the magnetic beads 260 are stopped or prevented from moving during subsequent steps of removing and adding fluid to the container 240.

Next, the purification method may include aspirating or removing the culture medium 242 from the container 240 via the fluid transfer member 214. To this end, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 horizontally, e.g., in the x-direction and/or the y-direction, until the fluid conduit 264a is disposed directly vertically above a first one of the containers 240 and the fluid conduit 264b is disposed directly vertically above a second one of the containers 240. Subsequently, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a downward vertical direction (i.e., along the z-axis) such that the first fluid conduit 264a is inserted through the opening 244 on the top of the first one of the containers 240 and is submerged in the media 242 of that container 240, and the second fluid conduit 264b is inserted through the opening 244 on the top of the second one of the containers 240 and is submerged in the media 242 contained by that container 240. This fluid transfer member 214 may be moved by the robotic manipulator 216 in a downward vertical direction until the mouth of the first fluid conduit 264a is adjacent or otherwise in close proximity to the bottom wall 250 of the first one of the containers 240 and the second fluid conduit 264b is adjacent or otherwise in close proximity to the bottom wall 250 of the second one of the containers 240. So configured, the first and second fluid conduits 264a, 264b can be capable of aspirating all or substantially all of the culture media 242 from their respective containers 240. Pumps 222a and 222b may be activated by control unit 230 after proper positioning of fluid transfer member 214 to generate the suction required to remove media 242 from container 240. The magnetic beads 260 are not removed during this step because the magnetic pull of the magnetic field generating elements 218a-e statically hold the beads against the inner surface 246 of the sidewall 248 of the container 240. Also, during this pumping step, the multi-position valve 224 may be controlled to fluidly connect each pump 222a and 222b with a waste container or drain 226 to expel the pumped fluid therefrom.

Once the aspiration is complete, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a vertical direction to remove the first and second fluid conduits 264a and 264b from their respective containers 240, and then the process in the preceding paragraph may be repeated for other containers 240 disposed on the work surface 212.

Next, a washing fluid (e.g., saline solution) may be added to the container 240 by the fluid transfer member 214 to clean the inner surface 246 of the container 240 and/or the magnetic beads 260 of any residual media 242 without removing the target species T from the magnetic beads 260. Initially, similar to the media removal step described above, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 horizontally such that the fluid conduit 264a is disposed directly vertically above a first one of the containers 240 and the fluid conduit 264b is disposed directly vertically above a second one of the containers 240. Subsequently, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a downward vertical direction (i.e., along the z-axis) such that the first fluid conduit 264a is inserted through the opening 244 on top of the first one of the containers 240 and the second fluid conduit 264b is inserted through the opening 244 on top of the second one of the containers 240. At or about the same time, the control unit 230 may control the multi-position valve 224 to fluidly connect each pump 222a and 222b with one or more auxiliary containers 228a-c containing wash fluid. Subsequently, the control unit 230 may activate the pumps 222a and 222b to draw the washing fluid from the one or more auxiliary containers 228a-c and transfer it to the container 240 via the fluid transfer member 214.

Once the wash fluid has been added to the first and second ones of the containers 240, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a vertical direction to remove the first and second fluid conduits 264a and 264b from their respective containers 240, and then the process in the preceding paragraph may be repeated for other containers 240 disposed on the work surface 212.

Alternatively, the magnetic field generating elements 218a-e may be translated from their respective operative positions adjacent the receptacle 240 to their respective inoperative positions away from the receptacle 240, either immediately prior to or after the addition of the washing fluid to the receptacle 240. In the absence of the relatively strong magnetic field of the magnetic field generating elements 218a-e, the magnetic beads 260 may be freely dispersed in the washing fluid, which may improve the efficiency of the washing process and/or allow for agitation or other agitation of the magnetic beads 260 in the washing fluid. In some embodiments, moving the magnetic field generating elements 218a-e may be accomplished by the control unit 230 activating the linear actuators 220a-e such that the linear actuators 220a-e each translate a respective one of the magnetic field generating elements 218a-e from its respective operative position to a respective inoperative position.

After washing is complete, the control unit 230 may control the multi-position valve 224 to fluidly connect the pumps 222a and 222b to the waste container or drain 226, and also control the robotic manipulator 216 to move the fluid transfer member 214 between different containers 240 while controlling the pumps 222a and 222b to draw wash fluid from the containers 240 and drain the wash fluid to the drain 226. Prior to this wash fluid removal step, the magnetic field generating elements 218a-e may be transferred from their respective inoperative positions away from the receptacle 240 to their respective operative positions adjacent to the receptacle 240, if not already completed. As a result, the magnetic field of the magnetic field generating elements 218a-e may statically attract and hold the magnetic beads 260 against the inner surface 246 of the sidewall 248 of their respective receptacle 240 while removing the washing fluid from the receptacle 240. The control unit 230 may be responsible for activating the linear actuators 220a-e to move the magnetic field generating elements 218a-e from their respective inoperative positions to their respective operative positions.

In alternative embodiments, one or more or all of the steps associated with the above-described wash fluid may be omitted.

Next, an eluent (e.g., a liquid elution solution) may be added to the container 240 by the fluid transfer component 214 to cause the target substance T to be released from the magnetic beads 260. As an initial step herein, the control unit 230 may control the multi-position valve 224 to fluidly connect the pumps 222a and 222b to one or more auxiliary containers 228a-c containing elution fluid. Also, if not completed, the magnetic field generating elements 218a-e may be translated from their respective operative positions adjacent to the receptacle 240 to their respective inoperative positions away from the receptacle 240, either just prior to or just after the addition of elution fluid to the receptacle 240. This may be accomplished by the control unit 230 activating the linear actuators 220a-e to move the magnetic field generating elements 218a-e from their respective inoperative positions to their respective inoperative positions. Then, similar to the wash fluid step described above, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 horizontally such that the fluid conduit 264a is disposed directly vertically above a first one of the containers 240 and the fluid conduit 264b is disposed directly vertically above a second one of the containers 240. Subsequently, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a downward vertical direction (i.e., along the z-axis) such that the first fluid conduit 264a is inserted through the opening 244 on top of the first one of the containers 240 and the second fluid conduit 264b is inserted through the opening 244 on top of the second one of the containers 240. Subsequently, the control unit 230 may activate the pumps 222a and 222b to draw the washing fluid from the one or more auxiliary containers 228a-c and transfer it to the container 240 via the fluid transfer member 214.

In the absence of the relatively strong magnetic field of the magnetic field generating elements 218a-e, the magnetic beads 260 may be freely dispersed in the elution fluid, which may increase the efficiency of the elution process and/or allow for stirring or other agitation of the magnetic beads 260 in the elution fluid. The eluent addition process described above can be repeated for all of the containers 240. In an alternative embodiment, this elution step may be performed with the magnetic field generating elements 218a-e arranged in their respective working positions.

After a period of incubation, the eluent and target species T may be removed from container 240 via fluid transfer member 214 and subsequently discharged into an external container. As an initial step herein, the control unit 230 may control the multi-position valve 224 to fluidly connect the pumps 222a and 222b to one or more auxiliary containers 228a-c for storing elution fluid mixed with the target substance T. Also, if not yet completed, the magnetic field generating elements 218a-e may be translated from their respective inoperative positions away from the container 240 to their respective operative positions adjacent to the container 240. This may be accomplished by the control unit 230 activating the linear actuators 220a-e to move the magnetic field generating elements 218a-e from their respective operative positions to their respective inoperative positions. As a result, the magnetic field of the magnetic field generating elements 218a-e may statically attract and hold the magnetic beads 260 against the inner surface 246 of the sidewall 248 of their respective container 240. Then, similar to the wash fluid step described above, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 horizontally such that the fluid conduit 264a is disposed directly vertically above a first one of the containers 240 and the fluid conduit 264b is disposed directly vertically above a second one of the containers 240. Subsequently, the control unit 230 may control the robotic manipulator 216 to move the fluid transfer member 214 in a downward vertical direction (i.e., along the z-axis) such that the first fluid conduit 264a is inserted through the opening 244 on top of the first one of the containers 240 and the second fluid conduit 264b is inserted through the opening 244 on top of the second one of the containers 240. Subsequently, the control unit 230 may activate the pumps 222a and 222b to draw elution fluid mixed with the target substance T from the container 240 and transfer it to one or more auxiliary containers 228 a-c. The process of removing the eluent mixed with the target substance T may be repeated for all the containers 240.

It is noted that any of the foregoing steps of moving the magnetic field generating element into and out of the operative and/or inoperative position may be performed manually by a user, such as a laboratory technician.

While the apparatus, systems, and methods of the present disclosure have been described in connection with various embodiments, it will be understood that the apparatus, systems, and methods of the present disclosure are capable of further modifications. This application is intended to cover any variations, uses, or adaptations of the devices, systems, and methods following, in general, the principles of the disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains.

Additionally, it should be noted that the construction and arrangement of the disclosed magnetic separators and workstations, as well as the various components and assemblies thereof, as shown in the various exemplary embodiments, are illustrative only. Although only a few embodiments of the subject matter and problems have been described in detail in this disclosure, those skilled in the art who review this disclosure will readily appreciate that many modifications are possible (e.g., variations in sizes, dimensions, structures, shapes and proportions of the various elements, values of parameters, mounting arrangements, use of materials, colors, orientations, etc.) without materially departing from the novel teachings and advantages of the subject matter disclosed herein. For example, elements shown as integrally formed may be constructed of multiple parts or elements, and vice versa. Also, the position of elements may be reversed or otherwise varied, and the nature or number of discrete elements or positions may be altered or varied. Accordingly, all such modifications are intended to be included within the scope of this disclosure as defined in the following claims. Additionally, the order or sequence of any process or method steps may be varied or re-sequenced according to alternative embodiments. Other substitutions, modifications, changes and omissions may be made in the design, operating conditions and arrangement of the various exemplary embodiments without departing from the scope of the present disclosure.