Magnetic assisted separation device and related methods

Abstract

Magnetic-assisted separation devices and related methods for separating a target substance from a culture medium in which the target substance is suspended are provided. According to one aspect, a magnetic separator can include a frame having an opening configured to receive one or more containers containing the culture medium. Further, the magnetic separator can include first and second magnetic field generating elements mounted on opposite sides of the frame such that the one or more containers can be positioned between the first and second magnetic field generating elements. According to another aspect, a workstation includes a work surface for receiving one or more containers containing the culture medium, a fluid transfer member, an automated manipulator configured to move the fluid transfer member, and a plurality of magnetic field generating elements each movable between a first position away from the one or more containers and a second position adjacent to the one or more containers.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 62 / 534,563, filed July 19, 2017, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure generally relates to separation apparatus and methods, and more specifically to the separation of target substances (such as biomolecules) from fluid culture media in which target substances are suspended, in order to facilitate downstream processing or analysis of the target substances. Background Technology

[0004] Certain diagnostic, research, and pharmaceutical manufacturing activities benefit from or require the isolation of target substances (such as proteins) contained in cell cultures or other biological mixtures. This task has been accomplished in the past using various techniques. Some of these techniques require altering the solubility of the target substance to cause it to precipitate from the biological mixture. Other techniques require centrifugation, where particles of different densities are separated by rotating them at high speed around a fixed point. Still others are based on chromatography, which requires passing the biological mixture through a filter medium in which the components of the mixture move at different rates.

[0005] Such conventional purification techniques, and others, are often time-consuming, labor-intensive, and / or limited to relatively small sample sizes. For example, centrifugation is typically performed in test tubes or bottles, which can limit the amount of material that can be processed in a given time. Moreover, some conventional purification techniques may require laboratory technicians to manually pipette fluids between various containers, which can be inefficient and may increase the risk of cross-contamination.

[0006] This disclosure describes apparatus and methods related to purification that provide advantageous alternatives to existing purification apparatus and methods and can address one or more of the challenges or needs mentioned herein, as well as provide additional benefits and advantages. Summary of the Invention

[0007] One aspect of this disclosure provides a magnetic separator for separating or removing a target substance from a culture medium in which a target substance is suspended. The magnetic separator may include a frame having a first opening configured to receive at least one container or dish containing the culture medium. The magnetic separator may further include a first magnetic field generating element and a second magnetic field generating element. The first and second magnetic field generating elements may be mounted at a distance from each other on opposite sides of the frame, such that the at least one container can be positioned between the first and second magnetic field generating elements.

[0008] Another aspect of this disclosure provides a purification method that may include: (a) adding a culture medium in which a target substance is suspended to a container or vessel; (b) adding a plurality of magnetic beads to the container, wherein the target substance is temporarily bound 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, wherein at least one of the first magnetic field generating element or the second magnetic field generating element magnetically attracts the plurality of magnetic beads and holds them against the inner surface of the container.

[0009] An additional aspect of this disclosure provides a workstation for separating a target substance from a culture medium in which a target substance is suspended. The workstation may include: a working surface for receiving at least one container or dish containing the culture medium; and a fluid transfer member configured to transfer fluid to and from the at least one container. Furthermore, the workstation may include: an automanipulator configured to move the fluid transfer member relative to the working surface. Moreover, the workstation may include a plurality of magnetic field generating elements, each movable relative to the working surface between a first position remote from the at least one container and a second position adjacent to the at least one container.

[0010] Another aspect of this disclosure provides a purification method, which may include: (a) providing a workstation having a working surface, an automanipulator, and a plurality of magnetic field generating elements, the automanipulator being movable relative to the working surface and carrying a fluid transfer member, the plurality of magnetic field generating elements being laterally spaced apart to define a plurality of rows; (b) adding at least one culture medium in which a target substance is suspended to a plurality of containers or dishes; (c) adding a plurality of magnetic beads to the plurality of containers, the target substance being temporarily bound to the plurality of magnetic beads; and (d) 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 the inner surface of a respective container in the plurality of containers. Attached Figure Description

[0011] This disclosure will be more fully understood from the following description taken in conjunction with the accompanying drawings. To illustrate other elements more clearly, some drawings may be simplified by omitting selected elements. In some drawings, the omission of these elements does not necessarily indicate the presence or absence of a particular element in any embodiment, unless explicitly stated in the corresponding written description. Furthermore, not all drawings need to be drawn to scale.

[0012] Figure 1 This is a top perspective view of an embodiment of a magnetic separator based on the principles of this disclosure.

[0013] Figure 2 yes Figure 1 The front perspective view of the magnetic separator shown.

[0014] Figure 3 yes Figure 1 The rear perspective view of the magnetic separator shown.

[0015] Figure 4 yes Figure 1 The illustrated cross-sectional view shows a magnetic separator containing a container holding a fluid culture medium and magnetic beads.

[0016] Figure 5 This is a perspective side view of another embodiment of a magnetic separator based on the principles of this disclosure.

[0017] Figures 6A to 6G The steps of one embodiment of the purification method according to the principles of this disclosure are illustrated schematically in chronological order.

[0018] Figure 7 This is a schematic front plan view of an embodiment of a workstation for magnetic separation based on the principles of this disclosure.

[0019] Figure 8 yes Figure 7 The workstation shown is represented schematically in a side plan view.

[0020] Figure 9 This is a front perspective view of an embodiment of a workstation based on the principles of this disclosure. Detailed Implementation

[0021] This disclosure generally relates to apparatus and methods for separating a target substance, such as a protein, from a fluid culture medium in which a target substance is suspended. In a broader sense, the apparatus and methods disclosed herein include positioning one or more magnetic field generating elements adjacent to the exterior of one or more containers containing a culture medium with the target substance. A plurality of magnetic beads may be immersed in the culture 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 culture medium can then be removed from the one or more containers, leaving the magnetic beads bound to the target substance. Subsequently, the magnetic beads may be subjected to washing and / or elution procedures to release and / or extract the target substance from the magnetic beads. The magnetic beads can ultimately be separated from the magnetic field and vice versa by simply removing the one or more magnetic field generating elements from the one or more containers.

[0022] This configuration advantageously simplifies the process of separating target substances from culture media, and in some embodiments, can eliminate or reduce the number of tasks that must be performed manually by laboratory technicians or other users. Furthermore, the currently disclosed apparatus and method can provide the capacity to process relatively large sample volumes without significantly increasing the workstation footprint required for purification-related equipment. In addition, the purification apparatus and method disclosed herein can reduce the possibility of cross-contamination, which can occur if one or more magnetic field generating elements are immersed in the sample culture medium.

[0023] Each of the aforementioned components and related methods will now be described in more detail.

[0024] The apparatus and methods disclosed herein can be used to isolate a wide variety of target substances (e.g., molecules, molecular complexes, biomolecules, biomolecular complexes, proteins, protein complexes, peptides, nucleic acid ligands, pathogenic microorganisms, cells, etc.) from a wide variety of sample mixtures (e.g., cell cultures, blood, tanshinone, mucus, sweat, urine, feces, soil, food, etc.). Moreover, depending on the characteristics of the target substance to be isolated and / or the culture medium in which the target substance is suspended, a wide variety of magnetic beads can be used. In some embodiments, the magnetic beads may have a spherical shape and a silica-based paramagnetic core coated with a material that binds to or is conjugated with the target substance. The binding between the magnetic beads and the target substance can be achieved covalently, non-covalently, or electrostatically via hydrogen bonding, van der Waals forces, and / or any other suitable molecular binding process. In at least one embodiment, the culture medium may be a cell culture or other biological mixture, the target substance may be an antibody or other protein, and the magnetic beads may be protein A magnetic beads. In another embodiment, the culture medium may be a cell culture or other biological mixture, the target substance may be a polyhistidine-labeled protein, and the magnetic beads may be coated with nickel, zinc, copper, or cobalt.

[0025] A non-limiting list of examples of magnetic bead types that can be implemented in the currently 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 resin magnetic beads, hydrazide magnetic beads, hydroxyl magnetic beads, iodoacetyl magnetic beads, NHS magnetic beads, mercapto magnetic beads, toluenesulfonyl magnetic beads, thiol magnetic beads, silica magnetic beads, IDA magnetic beads, etc.); reverse-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 magnetic beads (e.g., DEAE magnetic beads, PSA magnetic beads, SAX magnetic beads, WCX magnetic beads, SCX magnetic beads, hydroxyapatite magnetic beads, etc.); antibody purification magnetic beads (e.g., protein A magnetic beads, protein G magnetic beads, protein A / G magnetic beads, protein L magnetic beads, rapid IgG). Pure magnetic beads, antigen peptide magnetic beads, rapid IgM pure magnetic beads, anti-IgG magnetic beads, rapid IgA pure magnetic beads, thiophilic magnetic beads, etc.; antibody immobilization 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 magnetic beads (e.g., Ni+ banded electromagnetic beads, Co+ banded electromagnetic beads, maltose magnetic beads, calmodulin magnetic beads, etc.); peptide immobilization 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 endotoxins; magnetic beads for removing large amounts of protein; and / or EDTA magnetic beads.

[0026] As used herein, the term "magnetic" is defined to encompass any element that is magnetic, paramagnetic, and / or ferromagnetic. Therefore, a magnetic bead can be a magnetic bead, a paramagnetic bead, a ferromagnetic bead, or any combination thereof.

[0027] In some embodiments, the density of the magnetic beads may be greater than the density of the culture medium, such that when the magnetic beads are immersed in the sample mixture, they sink to the bottom of the container holding the culture medium. In other embodiments, the density of the magnetic beads may be less than or equal to the density of the culture medium, such that the magnetic beads float or partially float in the culture medium.

[0028] Figures 1 to 3 An embodiment of a magnetic separator 10 according to the principles of this disclosure is illustrated. The magnetic separator 10 typically includes a frame 12 for mounting magnetic field generating elements 14 and 16 spaced apart by a distance X1. An opening 18 may be formed in the frame 12, and its size allows one or more containers 20 to be inserted through the opening 18 into the space between the magnetic field generating elements 14 and 16, such as... Figure 3 As shown. In some embodiments, container 20 may be attached to non-magnetic support frame 50 (see...). Figure 4The non-magnetic support frame can also be positioned between the magnetic field generating elements 14 and 16, or held by it.

[0029] Typically, although there are magnetic attraction and / or repulsion between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the container 20, the frame 12 serves to maintain the spacing distance X1 between the magnetic field generating elements 14 and 16. Fixing the magnetic field generating elements 14 and 16 at the set distance X1 via the frame 12 may eliminate the need for the user to set or otherwise handle the magnetic field generating elements 14 and 16, which could be cumbersome and potentially unsafe when the magnetic pull between the magnetic field generating elements 14 and 16 is relatively strong. Furthermore, by avoiding direct contact with the magnetic field generating elements 14 and 16, the risk of damage to the memory or other sensitive components of the user-carried instruments or personal electronic devices is reduced.

[0030] In some embodiments, the lateral distance X1 separating the first magnetic field generating element 14 and the second magnetic field generating element 16 may be in the range of approximately (e.g., ±10%) 3 to 36 inches, or in the range of approximately (e.g., ±10%) 3 to 24 inches, or in the range of approximately (e.g., ±10%) 3 to 18 inches, or in the range of approximately (e.g., ±10%) 3 to 12 inches, or greater than or equal to approximately (e.g., ±10%) 1 inch, or greater than or equal to approximately (e.g., ±10%) 2 inches, or greater than or equal to approximately (e.g., ±10%) 3 inches, or greater than or equal to approximately (e.g., ±10%) 4 inches, or equal to approximately (e.g., ±10%) 3.4 inches.

[0031] In this embodiment, the spacing X1 between the magnetic field generating elements 14 and 16 is not adjustable. However, in an alternative embodiment, the frame 12 may have an adjustable width, thereby allowing the user to adjust the spacing X1 between the magnetic field generating elements 14 and 16, and then lock the magnetic field generating elements 14 and 16 into place.

[0032] Reference Figures 1 to 4 The frame 12 may be composed of a first vertical sidewall 30, a second vertical sidewall 32, and a horizontal bottom wall 34. The first vertical sidewall 30 and the second vertical sidewall 32 may be arranged parallel to each other as shown, or in an alternative embodiment, they may be arranged at a non-parallel angle relative to each other. The horizontal bottom wall 34 may extend between the first vertical sidewall 30 and the second vertical sidewall 32 and may provide the necessary structural support to prevent the first vertical sidewall 30 and the second vertical sidewall 32 from moving relative to each other due to the magnetic attraction and / or repulsion forces between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the container 20. Figures 1 to 3As shown, additional structural support can be provided by one or more beams 34a-d extending between the first vertical sidewall 30 and the second vertical sidewall 32. In an alternative embodiment, the bottom wall 34 can be omitted, and only one or more beams 34a-d can be included to keep the first vertical sidewall 30 and the second vertical sidewall 32 separated. In a further alternative embodiment, the beams 34a-d can be omitted, and only the bottom wall 34 can be included to maintain the spacing between the first vertical sidewall 30 and the second vertical sidewall 32.

[0033] like Figures 1 to 3 As depicted, frame 12 does not include a front wall, top wall, or rear wall. Instead of a front wall, an opening 18 (i.e., a front opening) is defined between a first vertical side wall 30 and a second vertical side wall 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 vertical side wall 30 and the second vertical side wall 32, respectively. In alternative embodiments, a top wall and / or a rear wall may be included, such that the openings are only the front opening 18 and the rear opening 28, or only the front opening 18 and the top opening 26, or only the front opening 18. In a further alternative embodiment, the top opening 26 may be the only opening formed in frame 12. In the illustrated embodiment, the front opening 18 is separated from the top opening 26 by beam 34a, and the top opening 26 is separated from the rear opening 28 by beam 34b. In other embodiments, with beams 34a and 34b omitted, the front opening 18, the top opening 26, and the rear opening 28 may be continuous with each other (e.g., see...). Figure 5 ).

[0034] Moreover, although the vertical sidewalls 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 by one or more pillars or trusses with gaps therebetween, or have one or more cut-out segments to reduce the weight of the frame 12.

[0035] The frame 12 may be made of a rigid material (including, but not limited to, certain types of metals and / or plastics). The rigidity of the frame 12 should be such that the magnetic attraction and / or repulsion forces between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the container 20 will not cause deformation of the frame 12. In some embodiments, the frame 12 may be made of 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.

[0036] Continue to refer to Figures 1 to 4The first magnetic field generating element 14 can be rigidly fixed to the inward-facing surface 36 of the first vertical sidewall 30, and the second magnetic field generating element 16 can be rigidly fixed to the 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 can be rigidly fixed, for example, with fasteners such as bolts and / or screws. Additionally, in some embodiments, each of the first magnetic field generating element 14 and / or the second magnetic field generating element 16 can be housed in a corresponding cage or other enclosure 44 or 46, which is rigidly fixed to the inward-facing surface 36 of the first vertical sidewall 30 or the inward-facing surface 38 of the second vertical sidewall 32. Each cage 44 and 46 may have a side opening 48 or 50, thereby allowing the first magnetic field generating element 14 or the second magnetic field generating element 16 to be slidably inserted into or removed from its corresponding cage 44 or 46.

[0037] Each of the magnetic field generating elements 14 and 16 can be mounted such that its downward-facing surface or bottom surface is spaced vertically from the horizontal bottom wall 34, or, if the horizontal bottom wall 34 is omitted, spaced a distance X2 from the surface on which the frame 12 rests. As discussed below, this can provide space or clearance for the sides of the non-magnetic support frames inserted below the first magnetic field generating element 14 and the second magnetic field generating element 16, respectively.

[0038] like Figure 4 As shown, the first magnetic field generating element 14 and the second magnetic field generating element 16 protrude inward from the inward-facing surface 36 of the first vertical sidewall 30 and the inward-facing surface 38 of the second vertical sidewall 32, respectively. In an alternative embodiment, the first magnetic field generating element 14 and the second magnetic field generating element 16 may be positioned in corresponding grooves or recesses formed in the inward-facing surfaces 36 of the first vertical sidewall 30 and 38 of the second vertical sidewall 32, respectively. Thus, the first magnetic field generating element 14 and the second magnetic field generating element 16 can be flush with the inward-facing surfaces 36 and 38, respectively.

[0039] In some embodiments, such as Figure 4 As shown, the first magnetic field generating element 14 and the second magnetic field generating element 16 can have a generally flat or planar shape with a rectangular cross-section. However, other shapes and cross-sections are also possible. Furthermore, magnetic field generating elements other than those depicted in the figures may 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 has no magnetic field generating element.

[0040] In some embodiments, each of the magnetic field generating elements 14 and 16 may be composed of a corresponding permanent magnet configured to generate its own persistent magnetic field. The maximum magnetic pull of each permanent magnet may be in the range of approximately (e.g., ±10%) 50 to 1000 Newtons (N), or approximately (e.g., ±10%) 100 to 800 N, or approximately (e.g., ±10%) 100 to 700 N, or approximately (e.g., ±10%) 150 to 600 N, or approximately (e.g., ±10%) 200 to 500 N, or approximately (e.g., ±10%) 200 to 450 N, or greater than or equal to approximately (e.g., ±10%) 50 N, or greater than or equal to approximately (e.g., ±10%) 100 N, or greater than or equal to approximately (e.g., ±10%) 150 N, or greater than or equal to approximately (e.g., ±10%) 200 N, or greater than or equal to approximately (e.g., ±10%) 250 N. In some embodiments, the total combined magnetic pull of the permanent magnets may be greater than or equal to about (e.g., ±10%) 500 N, or greater than or equal to about (e.g., ±10%) 1000 N, or greater than or equal to about (e.g., ±10%) 1500 N, or greater than or equal to about (e.g., ±10%) 2000 N, or greater than or equal to about (e.g., ±10%) 2500 N. In some embodiments, the permanent magnets constituting the magnetic field generating elements 14 and 16 may be nickel-plated neodymium block magnets. In alternative embodiments, each of the magnetic field generating elements 14 and 16 may be constituted by a corresponding electromagnet configured to generate a magnetic field when supplied with current.

[0041] Reference Figure 4 The magnetic separator 10 may include a nonmagnetic support frame 50, which is separate from and movable relative to the frame 12. Typically, the nonmagnetic support frame 50 may be configured to hold a plurality of containers 20 containing culture medium 54, in which the target substance T is initially suspended. In some embodiments, the nonmagnetic support frame 50 may be configured to prevent lateral movement of the containers 20 relative to each other due to magnetic attraction and / or repulsion between the magnetic field generating elements 14 and 16 and the magnetic beads 40 disposed in the containers 20. This aspect of the nonmagnetic support frame 50 may be achieved through various constructions, including, for example: forming a nonmagnetic support frame 50 with a plurality of holes, each sized to receive a container 20; or rigidly securing the containers 20 to the nonmagnetic support frame 50 by one or more fasteners or adhesives; or, as... Figure 5As shown in the illustrated embodiment, the container 120 and the support frame 150 are integrally joined together, forming a single, integral structure. The non-magnetic support frame 50 can be made of any non-magnetic material, including, for example, plastic or glass, so that it is not magnetically attracted or repelled by the first magnetic field generating element 14 and the second magnetic field generating element 16. The container 20 can also be made of a non-magnetic material, including, for example, glass or plastic.

[0042] In some embodiments, the non-magnetic support frame 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 by the non-magnetic support frame 50 may extend longitudinally in a direction parallel to the first vertical sidewall 30 and the second vertical sidewall 32.

[0043] The nonmagnetic support frame 50 may have a width or other dimensions configured such that it can be inserted in a generally horizontal direction through an opening 18 in the front side of the frame 12. This insertion causes the container 20 to be positioned in the space between the first magnetic field generating element 14 and the second magnetic field generating element 16. In some embodiments, such as... Figure 4 In the illustrated embodiment, the width W1 of the non-magnetic support frame 50 can be greater than the distance X1 separating the first magnetic field generating element 14 and the second magnetic field generating element 16. In such an embodiment, the height H1 of the non-magnetic support frame 50 can be less than the vertical distance X2 separating the bottom surfaces of the first magnetic field generating element 14 and the second magnetic field generating element 16 from the horizontal bottom wall 34 of the frame 12. Therefore, the first lateral end 56 and the second lateral end 58 of the non-magnetic support frame 50 can be respectively mounted below the first magnetic field generating element 14 and the second magnetic field generating element 16. This configuration can advantageously limit the lateral movement of the non-magnetic support frame 50, which is 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 containers 20. This is because the first lateral end 56 and the second lateral end 58 of the non-magnetic support frame 50 can respectively abut against the inward-facing surface 36 of the first vertical sidewall 30 and the inward-facing surface 38 of the first vertical sidewall 32, thereby preventing significant lateral movement of the non-magnetic support frame 50.

[0044] Go to Figure 5 The illustration shows another embodiment of a magnetic separator 110 according to the principles of this disclosure. The magnetic separator 110 is similar to the magnetic separator 10, except that it does not include a beam extending between the vertical walls 130 and 132 of the frame 112. Figure 5 The diagram and Figures 1 to 4 The elements of the magnetic separator 10 illustrated are the same as or similar to those of the magnetic separator 110, and are indicated by the same reference numerals increased by 100. For the sake of brevity, descriptions of these similar components are omitted.

[0045] The absence of beams extending between the vertical walls 130 and 132 of frame 112 makes the front opening 118, top opening 126, and rear opening 128 continuous with each other. Therefore, the space between the first magnetic field generating element 114 and the second magnetic field generating element 116 is more easily accessible to the user, and more than one insertion path for the non-magnetic support frame 150 is possible.

[0046] Now refer to Figures 6A to 6G Describes a method for using a magnetic separator 10 during the purification process. Figures 6A to 6G This is a schematic side view, and various components of the magnetic separator 10 are omitted. The omission of these components should not be construed as meaning they are necessarily missing from the magnetic separator 10. Furthermore, the magnetic separator 110 can use a similar... Figures 6A to 6G The process described in the text.

[0047] As an initial step, each container 20 or a single container 20 can be filled with a certain volume of culture medium 54 containing the target substance T. Next, as... Figure 6A As shown, magnetic beads 40 can be added to container 20 and allowed to interact with culture medium 54 for a period of time (e.g., minutes, hours, hours, days, etc.). During this incubation, magnetic beads 40 can bind to the target substance T, thereby separating the target substance T from the rest of the culture medium 54 (see [reference]). Figure 6B As previously described, the binding between magnetic beads 40 and the target substance T can be achieved covalently, non-covalently, or electrostatically through hydrogen bonding, van der Waals forces, and / or any other suitable molecular binding process. In some embodiments, the culture medium 54 can be stirred or agitated during incubation to promote the binding between the target substance T and the magnetic beads 40. Additionally, in some embodiments, the container 20 can be placed in the magnetic separator 10 during the period when the magnetic beads 40 bind to the target substance T.

[0048] Next, if not yet completed, the container 20 can be fixed to the non-magnetic support frame 50, and the non-magnetic support frame 50 can be inserted between the first vertical sidewall 30 and the second vertical sidewall 32 of the frame 12, thereby positioning the container 20 between the first magnetic field generating element 14 and the second magnetic field generating element 16, as follows. Figure 6CAs shown. This step may require the user to manually move the nonmagnetic support frame 50, on which the container 20 is mounted, from a first position outside the frame 12 to a second position between the first vertical sidewall 30 and the second vertical sidewall 32 of the frame 12. In some embodiments, this movement may include inserting the nonmagnetic support frame 50 and the container 20 horizontally through the front opening 18 of the frame 12. In some embodiments, the first and second lateral ends 56 and 58 of the nonmagnetic support frame 50 may slide or otherwise mate below the first magnetic field generating element 14 and the second magnetic field generating element 16 during insertion. In alternative embodiments, the nonmagnetic support frame 50 and the container 20 may be inserted vertically downward through the top opening 26 or horizontally through the rear opening 28.

[0049] like Figure 6C As shown, when 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 allows them to magnetically attract and hold the magnetic bead 40 against the inner surface 60 of their respective container 20 sidewall 61. The friction between the magnetic bead 40 and the inner surface 60 of container 20 effectively fixes or immobilizes the magnetic bead 40 relative to container 20, preventing or inhibiting movement of the magnetic bead 40 during subsequent fluid removal and addition steps.

[0050] Next, as Figure 6D As shown, the purification method may include aspirating or removing culture medium 54 from container 20 via a fluid transfer member 62, such as a pipette or other fluid conduit. The distal end or nozzle of the 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 the containers 20, thereby immersing it in the culture medium 54. The nozzle of the fluid transfer member 62 may be positioned adjacent to or otherwise very close to the bottom wall 66 of container 20, such that the fluid transfer member 62 can aspirate all or substantially all of the culture medium 54 in container 20. Magnetic beads 40 are not removed in this step because they are held against the inner surface 60 of the side wall 61 of container 20 by the magnetic pull of the first magnetic field generating element 14 and / or the second magnetic field generating element 16. Additionally, the target substance T bound to the magnetic beads 40 also remains in container 20. This step of removing culture medium 54 from which the target substance T has been removed may be repeated for all containers 20, or in some embodiments, this step may be performed simultaneously on all containers 20 using multiple fluid transfer members.

[0051] After removing the culture medium 54, a washing fluid 70 (e.g., a salt solution) can be added to the container 20 via the fluid transfer member 62 or another fluid conduit to clean the inner surface 60 of the container 20 and / or any residual culture medium 54 on the magnetic beads 40, such as... Figure 6EAs shown. However, the washing fluid 70 cannot remove the target substance T from the magnetic beads 40. The volume of the washing fluid 70 added to the container 20 can be equal to or less than the maximum volume of the corresponding container 20 (e.g., a small fraction of it). Optionally, just before or immediately after adding the washing fluid 70 to the container 20, the container 20, or the entire support frame of the container 20, can be removed from the frame 12, such that the container 20 is not positioned between the first magnetic field generating element 14 and the second magnetic field generating element 16. In the absence of the relatively strong magnetic fields of the magnetic field generating elements 14 and 16, the magnetic beads 40 can be freely dispersed in the washing fluid 60 and can be suspended in the washing fluid 60. Allowing the magnetic beads 40 to be dispersed in this way can improve the efficiency of the washing process and / or allow the magnetic beads 40 in the washing fluid 70 to be stirred or otherwise agitated. After washing is complete, the washing fluid 70 can be removed from the container 20 via the fluid transfer member 62 or other fluid conduit.

[0052] In alternative embodiments, one or more or all of the steps related to the washing fluid described above may be omitted.

[0053] Next, as Figure 6F As shown, eluent 72 (e.g., a liquid eluent solution) can be added to container 20 via fluid transfer component 62 or another fluid conduit to release the target substance T from magnetic beads 40. If not yet completed, container 20, or the entire support frame of container 20, is removed from frame 12 either before or immediately after adding eluent 72, so that container 20 is not positioned between the first magnetic field generating element 14 and the second magnetic field generating element 16. In the absence of the relatively strong magnetic fields of magnetic fields 14 and 16, magnetic beads 40 can be freely dispersed in eluent 72 and can be suspended in eluent 72. Allowing magnetic beads 40 to be dispersed in this manner can improve the efficiency of the elution process and / or allow for stirring or other agitation of magnetic beads 40 in eluent 72. Alternatively, this elution step can be performed while container 20 is positioned between the first magnetic field generating element 14 and the second magnetic field generating element 16.

[0054] After a period of incubation, the eluent 72 and the target substance T bound to the eluent 72 can be removed from container 20 via fluid transfer member 62 or another conduit, and then discharged into an outer container. If this is not yet complete, just before this fluid removal step, container 20 can be repositioned between the first magnetic field generating element 14 and the second magnetic field generating element 16 so that the magnetic beads 40 are fixed against the inner surface 60 of their respective containers 20. The above steps can then be repeated for another container or another batch of containers containing another culture medium or mixture that needs to be purified.

[0055] Note that any of the aforementioned steps of positioning container 20 between the first magnetic field generating element 14 and the second magnetic field generating element 16 and / or removing container 20 from between the first magnetic field generating element 14 and the second magnetic field generating element 16 can be performed manually by a user, such as a laboratory technician. Furthermore, any fluid addition or removal steps can be performed manually by the user using, for example, a pipette. Additionally, it should be noted that the magnetic separator 10 is sized and constructed such that it can be supported on various standard laboratory workbenches and / or tabletops.

[0056] Turn Figure 7 and Figure 8 The illustration shows a schematic representation of a workstation 200 for separating a target substance from a culture medium in which the target substance is suspended, according to the principles of this disclosure. Workstation 200 is similar in some respects to the magnetic separator embodiments described above in that during fluid removal steps, such as during removal of culture medium, removal of wash fluid, and / or removal of elution fluid, one or more magnetic field generating elements are used to immobilize magnetic beads immersed in the culture medium and bound to the target substance. However, workstation 200 differs from the aforementioned embodiments of the magnetic separator in that, while the container remains stationary, one or more magnetic field generating elements move between different locations during the washing and purification process. Additionally, unlike the magnetic separator described above, workstation 200 includes an automanipulator configured to move at least one fluid transfer member relative to the container for removing and / or adding fluid.

[0057] Typically, the workstation 200 is sized or otherwise constructed such that it can be supported on various standard laboratory workbenches and / or desktops. (See reference) Figures 7 to 9 Workstation 200 typically includes a frame 210 comprising a horizontally arranged, upward-facing work surface 212, a fluid transfer member 214, an automanipulator 216, and one or more magnetic field generating elements 218a-e. Optionally, 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 discharge port 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.

[0058] like Figure 7As shown, the magnetic field generating elements 218a-e can be laterally spaced from each other in a regular interval in the horizontal direction to define a plurality of rows 232a-d for slidably receiving a plurality of containers 240. In some embodiments, the lateral distance X3 between adjacent magnetic field generating elements in the magnetic field generating elements 218a-e can be in the range of approximately (e.g., ±10%) 3 to 10 inches, or in the range of approximately (e.g., ±10%) 3 to 8 inches, or in the range of approximately (e.g., ±10%) 3 to 6 inches, or in the range of approximately (e.g., ±10%) 3 to 5 inches, or greater than or equal to approximately (e.g., ±10%) 1 inch, or greater than or equal to approximately (e.g., ±10%) 2 inches, or greater than or equal to approximately (e.g., ±10%) 3 inches, or greater than or equal to approximately (e.g., ±10%) 4 inches, or equal to approximately (e.g., ±10%) 3.4 inches.

[0059] Each container 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 its top, an inner surface 246, a sidewall 248, and a bottom wall 250 connected to the sidewall 248. The culture medium 242 and the target substance T may be similar to the culture medium and target substance discussed above. Within each row 232a-d, the container 240 may be held by a non-magnetic support 252, which in some embodiments may be configured in a manner similar to the non-magnetic support discussed above.

[0060] Multiple magnetic beads 260 can be immersed in culture medium 242 in each container 240. The magnetic beads 260 can be constructed in a similar manner to the magnetic beads 40 described above, and can be configured to bind to the target substance T suspended in the culture medium 242.

[0061] Each magnetic field generating element 218a-e can extend longitudinally along the longitudinal axis A (see...). Figure 8Additionally, in some embodiments, the longitudinal axes A of the magnetic field generating elements 218a-e may be parallel to each other. The length L2 of each magnetic field generating element 218a-e may be parallel to the longitudinal axis A of the corresponding 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 corresponding magnetic field generating element. In some embodiments, the length L2 may be in the range of approximately (e.g., ±10%) 10 to 48 inches, or in the range of approximately (e.g., ±10%) 10 to 36 inches, or in the range of approximately (e.g., ±10%) 10 to 30 inches, or in the range of approximately (e.g., ±10%) 10 to 24 inches, or equal to or greater than approximately (e.g., ±10%) 10 inches, or equal to or greater than approximately (e.g., ±10%) 20 inches, or equal to or greater than approximately (e.g., ±10%) 30 inches. In some embodiments, the width W2 may be in the range of approximately (e.g., ±10%) 0.5 to 6 inches, or in the range of approximately (e.g., ±10%) 0.5 to 4 inches, or in the range of approximately (e.g., ±10%) 0.5 to 3 inches, or in the range of approximately (e.g., ±10%) 0.5 to 2 inches, or equal to or greater than approximately (e.g., ±10%) 0.5 inches, or equal to or greater than approximately (e.g., ±10%) 1 inch, or equal to or greater than approximately (e.g., ±10%) 1.5 inches, or equal to or greater than approximately (e.g., ±10%) 2 inches, or equal to or greater than approximately (e.g., ±10%) 2.5 inches.

[0062] Each magnetic field generating element 218a-e can move back and forth relative to the working surface 212 between a first or non-working position away from the corresponding row of containers 240 and a second or working position adjacent to (e.g., immediately adjacent to) the corresponding row of containers 240. Figure 8 One of the magnetic field generating elements 218a-e occupying its working position is schematically illustrated with solid lines. Figure 8The non-operating position of this magnetic field generating element among the magnetic field generating elements 218a-e is also illustrated by dashed lines. In this embodiment, the workstation 200 includes a plurality of linear actuators 220a-e, each configured to independently reciprocate a corresponding magnetic field generating element among the magnetic field generating elements 218a-e between its operating and non-operating positions in a linear horizontal direction. In other embodiments, a single linear actuator may be used to cause all magnetic field generating elements 218a-e to move simultaneously between their operating and non-operating positions. Each linear actuator 220a-e may include at least one of a hydraulic cylinder, a pneumatic cylinder, or an electric motor. In embodiments including a hydraulic cylinder or a pneumatic cylinder, a reciprocating piston may connect the hydraulic cylinder or pneumatic cylinder to a corresponding magnetic field generating element among the magnetic field generating elements 218a-e. In embodiments including a rotary electric motor, a rack and pinion mechanism, pulley, or gear system may be used to convert the rotary motion output generated by the electric motor into linear motion of the corresponding magnetic field generating element among the magnetic field generating elements 218a-e.

[0063] exist Figure 8 In the illustrated embodiment, each linear actuator 220a-e is in the form of a pneumatic linear slider. Thus, each linear actuator 220a-e may include a pneumatic cylinder 221a-e for receiving pressurized gas, and a piston or carrier element 223a-e that translates back and forth by pressurizing different ends of the pneumatic cylinder 221a-e. Figure 8 As shown, each pneumatic cylinder 221a-e can be arranged vertically below the working surface 212 within the interior space of the frame 210. Each bearing element 223a-e can be mechanically connected to a corresponding magnetic field generating element in the magnetic field generating element 218a-e through a corresponding opening 225a-e formed in the working surface 212. Although Figure 8 Only the linear actuator 220e and its connection to the magnetic field generating element 218e are illustrated; however, the other magnetic field elements 218a-d and the linear actuators 220a-d can be configured in a similar manner and are not illustrated for simplicity. In an alternative embodiment, the linear actuator 220e can be configured to move all the magnetic field generating elements 218a-e together.

[0064] Although the magnetic field generating elements 218a-e in this embodiment can move in a horizontal direction parallel to the working surface 212, in an alternative embodiment, each magnetic field generating element 218a-e can move in a vertical direction perpendicular to or otherwise not parallel to the working surface 212. In such an alternative embodiment, multiple openings can be formed in the working surface 212, and each magnetic field generating element 218a-e can be configured to reciprocately retract into and extend from a corresponding opening in these openings. Here, the retracted position (which may be below the working surface 212) may correspond to the non-working position, while the extended position (which may be above the working surface 212) may correspond to the working position.

[0065] In some embodiments, such as Figure 9 As illustrated, the linear actuators 220a-e can be omitted. In such an embodiment, the movement of the magnetic field generating elements 218a-e can be achieved by the user manually moving each magnetic field generating element 218a-e between its operating and non-operating positions in the horizontal and / or vertical directions.

[0066] 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 can effectively fix or immobilize the magnetic beads 260 relative to their respective containers 240, thereby preventing or stopping the movement of the magnetic beads 260 during subsequent fluid removal and / or addition steps.

[0067] In some embodiments, each magnetic field generating element 218a-e may be composed of a corresponding permanent magnet configured to generate its own persistent magnetic field. The maximum magnetic pull of each permanent magnet may be in the range of approximately (e.g., ±10%) 50 to 1000 Newtons (N), or in the range of approximately (e.g., ±10%) 100 to 800 N, or in the range of approximately (e.g., ±10%) 100 to 700 N, or in the range of approximately (e.g., ±10%) 150 to 600 N, or in the range of approximately (e.g., ±10%) 200 to 500 N, or in the range of approximately (e.g., ±10%) 2... The range is between 0 and 450 N, or between approximately (e.g., ±10%) 250 and 350 N, or greater than or equal to approximately (e.g., ±10%) 50 N, or greater than or equal to approximately (e.g., ±10%) 100 N, or greater than or equal to approximately (e.g., ±10%) 150 N, or greater than or equal to approximately (e.g., ±10%) 200 N, or greater than or equal to approximately (e.g., ±10%) 250 N, or equal to approximately (e.g., ±10%) 289 N. In some embodiments, the total combined magnetic pull of the permanent magnets may be greater than or equal to about (e.g., ±10%) 500 N, or greater than or equal to about (e.g., ±10%) 1000 N, or greater than or equal to about (e.g., ±10%) 1500 N, or greater than or equal to about (e.g., ±10%) 2000 N, or greater than or equal to about (e.g., ±10%) 2500 N, greater than or equal to about (e.g., ±10%) 5000 N, or greater than or equal to about (e.g., ±10%) 7000 N, or greater than or equal to about (e.g., ±10%) 7500 N, or equal to about (e.g., ±10%) 7225 N. In some embodiments, the permanent magnets constituting the magnetic field generating elements 218a-e may be nickel-plated neodymium block magnets. In alternative embodiments, each magnetic field generating element 218a-e may be constituted by a corresponding electromagnet configured to generate a magnetic field when supplied with current.

[0068] Continue to refer to Figures 7 to 9The automanipulator 216 can be configured to automatically move the fluid transfer member 214 relative to the working surface 212 between different positions, which may or may not be predefined or preprogrammed. The automanipulator 216 may include one or more electric motors controlled by the control unit 230. In the illustrated embodiment, the automanipulator 216 takes the form of a Cartesian coordinate robot capable of independent movement in each of the x, y, and z directions. In this embodiment, the x and y directions are horizontal linear directions, while the z direction is a vertical linear direction. The automanipulator 216 may include a separate electric motor and a separate track member for each direction of movement. In other embodiments, the automanipulator 216 may be capable of movement in only two directions (e.g., only x and y directions, or only x and z directions, or only y and z directions) or only in a single linear direction. In yet another embodiment, the automanipulator 216 may take the form of a robotic arm capable of performing compound bending, linear, and / or rotational movements.

[0069] Still refer to Figures 7 to 9 The autoactor 216 may include a mounting plate 262 for mounting the fluid transfer member 214, pumps 222a-b, multi-position valve 224, and / or other components to be carried by the autoactor 216. The fluid transfer member 214 may be mechanically connected (e.g., fastened with fasteners) to the mounting plate 262 and may extend downwards from the mounting plate 262 in a vertical direction. In this embodiment, the fluid transfer member 214 includes two parallel vertical straight-line conduits 264a and 264b (e.g., flexible or rigid plastic tubing) spaced horizontally from each other. Fluid conduit 264a may be in fluid communication with pump 222a, and fluid conduit 264b may be in fluid communication with pump 222b. The horizontal distance between the two fluid conduits 264a and 264b may be equal to the horizontal distance between the centers of any two adjacent rows 232a-d. Additionally, in some embodiments, the horizontal distance between the two fluid conduits 264a and 264b can be adjustable to accommodate containers 240 of different sizes. In alternative embodiments, the fluid transfer member 214 may comprise only a single vertical direct current conduit, or three or more vertical direct current conduits.

[0070] Typically, each pump 222a-b can be configured to remove and / or add fluid to a corresponding container in 240 via fluid transfer member 214. Each pump 222a-b can be driven by any suitable means, including but not limited to electric motors and / or pressurized hydraulic fluid and / or gas sources. Depending on the specifications of the purification process, each pump 222a-b can operate at a variable speed or a single speed. In some embodiments, the operation of one or more pumps 222a-b can be electronically controlled by control unit 230 according to programmable instructions, for example, stored in the memory of control unit 230. Alternatively or additionally, each pump 222a-b can be operated manually by a user (e.g., a laboratory technician) by actuating an ON / OFF switch and / or rotating a speed knob. In some embodiments, such as in Figure 9 In one embodiment depicted, one or more pumps 222a-b may be configured as positive displacement pumps (e.g., peristaltic pumps) and capable of pumping fluids containing suspended solids (e.g., magnetic beads 260) without damaging those solids. In alternative embodiments, one or more pumps 222a-b may be centrifugal pumps (e.g., radial pumps) that employ rotating impellers to create a vacuum to move the fluid. Additionally, in some embodiments, each pump 222a-b may be reversible.

[0071] Multi-position valve 224 can be configured to selectively connect pumps 222a and / or 222b fluidly to one or more of auxiliary containers 228a-c, discharge port 226, and / or other elements. Depending on the number of auxiliary containers, discharge ports, etc., multi-position valve 224 can be a 3-way valve, 4-way valve, 5-way valve, 6-way valve, 7-way valve, 8-way valve, or any other valve having any number of selectively openable orifices. In some embodiments, operation of multi-position valve 224 can be electronically controlled by control unit 230 according to programmable instructions, for example, stored in the memory of control unit 230. Multi-position valve 224 may be incorporated with one or more solenoids for opening and closing the orifices of multi-position valve 224 in response to command signals from control unit 230.

[0072] Typically, control unit 230 may be electrically connected at least to automanipulator 216, pumps 222a-b, and linear actuators 220a-e (if included), such that control unit 230 can send electrical control signals to and / or receive electrical control signals from these components. Control unit 230 may include a processor (e.g., a microprocessor), memory for storing tangible, non-transitory computer-readable instructions (e.g., random access memory (RAM), non-volatile memory (e.g., a processor-executable hard disk), flash memory, removable memory, non-removable memory, etc.), a communication unit, a display, and input devices (e.g., a keyboard, keypad, touchscreen, etc.). In some embodiments, control unit 230 may be a programmable logic controller. Control unit 230 can be programmed to perform a purification process according to user-defined specifications. In some embodiments, the control unit 230 may, in response to sensor data received from one or more sensors included in or used in conjunction with the workstation 200 and representing characteristics of the culture medium 242 and / or the target substance T (e.g., volume, temperature, weight, pH, etc.), timers, operator analog or digital inputs, and / or any other relevant detectable events or occurrences, perform steps of the purification process, such as activating the automanipulator 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.

[0073] The method of using workstation 200 during the purification process will now be described. As an initial step, each container 240 or a single container 240 can be filled with a certain volume of culture medium 242 containing the target substance T. Next, 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, several hours, a day, several days, etc.). During this incubation, the magnetic beads 260 can bind to the target substance T, thereby separating the target substance T from the remainder of the culture medium 242 (similar to...). Figure 6B As previously described, the binding between magnetic beads 260 and target substance T can be achieved covalently, non-covalently, or electrostatically through hydrogen bonding, van der Waals forces, and / or any other suitable molecular binding process. In some embodiments, the culture medium 242 can be stirred or agitated during incubation to promote the binding between target substance T and magnetic beads 242.

[0074] Next, if not yet completed, containers 240 can be grouped and secured to multiple non-magnetic support frames 252. Each loaded non-magnetic support frame 252 can then be horizontally inserted between a corresponding pair of magnetic field generating elements 218a-e on the top of the working surface 212. In doing so, each container 240 can be arranged in one of the rows 232a-d defined between adjacent magnetic field generating elements 218a-e. This step may include the user manually positioning each non-magnetic support frame 252 in its corresponding row of rows 232a-d. This insertion step can be performed with each magnetic field generating element 218a-e positioned in its respective working position (see [link to relevant documentation]). Figure 9 Alternatively, each magnetic field generating element 218a-e may be positioned in its respective working position. In the latter case, after the non-magnetic support 252 is placed on the working surface 212, the magnetic field generating elements 218a-e can be moved to their respective working positions.

[0075] The proximity of the magnetic field generating elements 218a-e allows them to magnetically attract the magnetic beads 260 and hold them against the inner surface 246 of the sidewall 248 of their respective containers 240. The friction between the magnetic beads 260 and the inner surface 246 of the container 240 effectively fixes or immobilizes the magnetic beads 260 relative to the container 240, thereby preventing or inhibiting movement of the magnetic beads 260 during subsequent steps of removing and adding fluid to the container 240.

[0076] Next, the purification method may include aspirating or removing culture medium 242 from container 240 via fluid transfer member 214. For this purpose, control unit 230 may control automanipulator 216 to move fluid transfer member 214 horizontally, for example, in the x and / or y directions, until fluid conduit 264a is positioned directly vertically above the first container in container 240, and fluid conduit 264b is positioned directly vertically above the second container in container 240. Subsequently, control unit 230 may control automanipulator 216 to move 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 at the top of the first container in container 240 and immersed in the culture medium 242 of container 240, and the second fluid conduit 264b is inserted through the opening 244 at the top of the second container in container 240 and immersed in the culture medium 242 contained in container 240. This fluid transfer member 214 can be moved vertically downwards by the automatic manipulator 216 until the nozzle of the first fluid conduit 264a is adjacent to or otherwise very close to the bottom wall 250 of the first container in container 240 and the second fluid conduit 264b is adjacent to or otherwise very close to the bottom wall 250 of the second container in container 240. With this configuration, the first fluid conduit 264a and the second fluid conduit 264b can aspirate all or substantially all of the culture medium 242 from their respective containers 240. Pumps 222a and 222b can be activated by the control unit 230 after the fluid transfer member 214 is correctly positioned to generate the suction required to remove the culture medium 242 from the containers 240. The magnetic beads 260 are not removed in this step because the magnetic pull of the magnetic field generating elements 218a-e statically holds the magnetic beads against the inner surface 246 of the sidewall 248 of the container 240. Furthermore, during this suction step, the multi-position valve 224 can be controlled to fluidly connect each pump 222a and 222b to the waste container or discharge port 226, thereby discharging the suctioned fluid there.

[0077] Once suction is complete, the control unit 230 can control the automanipulator 216 to move the fluid transfer members 214 in the vertical direction to remove the first and second fluid conduits 264a and 264b from their respective containers 240, and the process described in the preceding paragraph can then be repeated for other containers 240 disposed on the working surface 212.

[0078] Next, a washing fluid (e.g., a salt solution) can be added to container 240 via fluid transfer member 214 to clean the inner surface 246 of container 240 and / or any residual culture medium 242 on the magnetic beads 260 without removing the target substance T from the magnetic beads 260. Initially, similar to the culture medium removal step described above, control unit 230 can control automanipulator 216 to move fluid transfer member 214 horizontally, such that fluid conduit 264a is directly vertically positioned above the first container in container 240 and fluid conduit 264b is directly vertically positioned above the second container in container 240. Subsequently, control unit 230 can control automanipulator 216 to move 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 at the top of the first container in container 240, and the second fluid conduit 264b is inserted through the opening 244 at the top of the second container in container 240. Simultaneously or approximately simultaneously, control unit 230 may control multi-position valve 224 to fluidly connect each pump 222a and 222b to one or more auxiliary containers 228a-c containing washing fluid. Subsequently, control unit 230 may activate pumps 222a and 222b to draw washing fluid from one or more auxiliary containers 228a-c and transfer it to container 240 via fluid transfer member 214.

[0079] Once the washing fluid has been added to the first and second containers in container 240, the control unit 230 can control the automatic manipulator 216 to move the fluid transfer member 214 in the vertical direction to remove the first and second fluid conduits 264a and 264b from their respective containers 240, and the process in the preceding paragraph can then be repeated for the other containers 240 set on the working surface 212.

[0080] Optionally, just before or after the washing fluid is added to the container 240, the magnetic field generating elements 218a-e can be translated from their respective operating positions adjacent to the container 240 to their respective non-operating positions away from the container 240. In the absence of a relatively strong magnetic field from the magnetic field generating elements 218a-e, the magnetic beads 260 can be freely dispersed in the washing fluid, which can 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, the movement of the magnetic field generating elements 218a-e can be achieved by the control unit 230 activating the linear actuators 220a-e, such that each of the linear actuators 220a-e translates the respective magnetic field generating element of the magnetic field generating elements 218a-e from its respective operating position to its respective non-operating position.

[0081] After washing is complete, control unit 230 can control multi-position valve 224 to fluidly connect pumps 222a and 222b to waste container or discharge port 226, and also control autoacter 216 to move fluid transfer member 214 between different containers 240, while controlling pumps 222a and 222b to draw washing fluid from container 240 and discharge washing fluid to discharge port 226. Prior to this washing fluid removal step, if not yet completed, magnetic field generating elements 218a-e can be moved from their respective non-operating positions away from container 240 to their respective operating positions adjacent to container 240. As a result, the magnetic field of magnetic field generating elements 218a-e can statically attract and hold magnetic beads 260 against the inner surface 246 of the sidewall 248 of their respective containers 240, while removing washing fluid from container 240. Control unit 230 can be responsible for activating linear actuators 220a-e to move magnetic field generating elements 218a-e from their respective non-operating positions to their respective operating positions.

[0082] In alternative embodiments, one or more or all of the steps related to the washing fluid described above may be omitted.

[0083] Next, eluent (e.g., liquid eluent solution) can be added to container 240 via fluid transfer component 214 to release the target substance T from magnetic bead 260. As an initial step here, control unit 230 can control multi-position valve 224 to fluidly connect pumps 222a and 222b to one or more auxiliary containers 228a-c containing the eluent. Moreover, if not yet completed, either before or immediately after the addition of eluent to container 240, magnetic field generating elements 218a-e can be translated from their respective operating positions adjacent to container 240 to their respective non-operating positions away from container 240. This can be achieved by control unit 230 activating linear actuators 220a-e to move magnetic field generating elements 218a-e from their respective non-operating positions to their respective non-operating positions. Then, similar to the washing fluid steps described above, the control unit 230 can control the automatic actuator 216 to move the fluid transfer member 214 horizontally, such that the fluid conduit 264a is directly and vertically positioned above the first container in the container 240 and the fluid conduit 264b is directly and vertically positioned above the second container in the container 240. Subsequently, the control unit 230 can control the automatic actuator 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 at the top of the first container in the container 240, and the second fluid conduit 264b is inserted through the opening 244 at the top of the second container in the container 240. The control unit 230 can then activate pumps 222a and 222b to draw washing fluid from one or more auxiliary containers 228a-c and transfer it to the container 240 via the fluid transfer member 214.

[0084] In the absence of a relatively strong magnetic field from the magnetic field generating elements 218a-e, the magnetic beads 260 can be freely dispersed in the eluent, which can improve the efficiency of the elution process and / or allow for stirring or other agitation of the magnetic beads 260 in the eluent. The above-described eluent addition process can be repeated for all containers 240. In an alternative embodiment, this elution step can be performed with the magnetic field generating elements 218a-e arranged in their respective operating positions.

[0085] After a period of incubation, the eluent and target substance T can be removed from container 240 via fluid transfer member 214 and subsequently discharged into an external container. As an initial step here, control unit 230 can control multi-position valve 224 to fluidly connect pumps 222a and 222b to one or more auxiliary containers 228a-c for storing the eluent mixed with the target substance T. Moreover, if not yet completed, magnetic field generating elements 218a-e can be translated from their respective non-operating positions away from container 240 to their respective operating positions adjacent to container 240. This can be achieved by control unit 230 activating linear actuators 220a-e to move magnetic field generating elements 218a-e from their respective operating positions to their respective non-operating positions. As a result, the magnetic field of magnetic field generating elements 218a-e can statically attract and hold magnetic beads 260 against the inner surface 246 of the sidewall 248 of their respective containers 240. Then, similar to the washing fluid steps described above, the control unit 230 can control the automatic manipulator 216 to move the fluid transfer member 214 horizontally, such that the fluid conduit 264a is directly vertically positioned above the first container in container 240 and the fluid conduit 264b is directly vertically positioned above the second container in container 240. Subsequently, the control unit 230 can control the automatic 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 at the top of the first container in container 240, and the second fluid conduit 264b is inserted through the opening 244 at the top of the second container in container 240. The control unit 230 can then activate pumps 222a and 222b to draw the eluent mixed with the target substance T from container 240 and transfer it to one or more auxiliary containers 228a-c. The process of removing the eluent mixed with the target substance T can be repeated for all containers 240.

[0086] Note that any of the aforementioned steps of moving the magnetic field generating element into and out of the working and / or non-working position can be performed manually by a user, such as a laboratory technician.

[0087] While the apparatuses, systems, and methods disclosed herein have been described in conjunction with various embodiments, it will be understood that the apparatuses, systems, and methods disclosed herein can be further modified. This application is intended to cover any variations, uses, or modifications of the apparatuses, systems, and methods that generally follow the principles of this disclosure, and includes deviations from this disclosure within the scope of known and customary practices in the art to which this invention pertains.

[0088] Additionally, it should be noted that the construction and arrangement of the disclosed magnetic separators and workstations, as well as their various parts and components, are illustrative only, as shown in the various exemplary embodiments. Although only a few embodiments of the subject matter and problems are described in detail in this disclosure, those skilled in the art will readily understand that many modifications are possible (e.g., size, dimensions, structure, shape and proportion of different elements, parameter values, mounting arrangements, use of materials, color, orientation, etc.) without substantially departing from the novel teachings and advantages of the subject matter disclosed herein. For example, an element shown as integrally formed may be composed of multiple parts or elements, and vice versa. Moreover, the positions of elements may be reversed or otherwise changed, and the nature or number or position of discrete elements may be changed or varied. Therefore, all such modifications are intended to be included within the scope of this disclosure as defined by the appended claims. Furthermore, according to alternative embodiments, the order or sequence of any process or method steps may be changed or reordered. Other substitutions, modifications, alterations, and omissions may be made in the design, operating conditions, and arrangements of the various exemplary embodiments without departing from the scope of this disclosure.

Claims

1. A workstation for separating a target substance from a culture medium in which the target substance is suspended, the culture medium being contained in at least one container, the workstation comprising: A base, comprising a working surface for receiving the at least one container containing the culture medium, the working surface being arranged horizontally; A fluid transfer member configured to transfer fluid to and from the at least one container; An automatic actuator configured to move the fluid transfer member relative to the working surface; A peristaltic pump configured to add fluid to and remove fluid from the at least one container via the fluid transfer member; A valve having multiple positions, the valve being configured to connect the peristaltic pump to at least one external fluid source in a first position and to connect the peristaltic pump to at least one waste container or discharge pipe in a second position; Multiple magnetic field generating elements, each of which is capable of moving in the horizontal direction relative to the working surface between a first position away from the at least one container and a second position adjacent to the at least one container; as well as An actuator, at least partially vertically positioned within the base below the working surface, and configured to move at least a first magnetic field generating element among a plurality of magnetic field generating elements relative to the working surface. The fluid transfer component, peristaltic pump, and valve are mounted on the automatic actuator, such that the fluid transfer component, peristaltic pump, and valve move together during the movement of the automatic actuator.

2. The workstation of claim 1, wherein the plurality of magnetic field generating elements are laterally spaced apart to define a plurality of rows for receiving a plurality of containers, the plurality of containers comprising: At least one container.

3. The workstation as claimed in claim 2, wherein each of the plurality of magnetic field generating elements has a longitudinal axis, and the longitudinal axes of the plurality of magnetic field generating elements are parallel to each other.

4. The workstation as claimed in claim 3, wherein each of the plurality of magnetic field generating elements has 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.

5. The workstation as described in any one of claims 1 to 4, wherein, The actuator includes a linear actuator configured to cause at least the first magnetic field generating element of the plurality of magnetic field generating elements to reciprocate between the first position and the second position.

6. The workstation of claim 5, wherein the linear actuator comprises at least one of a hydraulic cylinder, a pneumatic cylinder, or an electric motor.

7. The workstation of claim 1, wherein the automatic manipulator is disposed above the work surface, and the fluid transfer member is configured to transfer fluid to and from the at least one container via an opening in the top of the at least one container.

8. The workstation of claim 1, wherein the automated manipulator comprises a Cartesian coordinate robot capable of movement in at least the x and y directions.

9. The workstation as described in claim 1, wherein, The valve is a multi-position valve, which is in fluid communication with the peristaltic pump.

10. The workstation of claim 1, comprising a plurality of magnetic beads for immersion in a culture medium in the at least one container, each of the plurality of magnetic beads having an outer surface configured to temporarily bind to the target substance to separate the target substance from the culture medium.

11. The workstation as claimed in claim 1, wherein, The fluid transfer component includes at least two parallel fluid conduits.

12. The workstation of claim 1, wherein the first magnetic field generating element comprises a first permanent magnet.

13. The workstation of claim 12, wherein the first permanent magnet has a maximum magnetic pull force equal to or greater than 100 N.

14. A purification method, comprising: A workstation is provided having a base with a horizontally arranged work surface, an automatic actuator, a valve, and a plurality of magnetic field generating elements. The automatic actuator is movable relative to the work surface and carries a fluid transfer member. A peristaltic pump is configured to add fluid to and remove fluid from a plurality of containers via the fluid transfer member. The valve has multiple positions and is configured to connect the peristaltic pump to at least one external fluid source in a first position and to connect the peristaltic pump to at least one waste container or discharge pipe in a second position. The plurality of magnetic field generating elements are laterally spaced apart from each other to define a plurality of rows. An actuator is at least partially vertically positioned below the work surface within the base. The fluid transfer member, the peristaltic pump, and the valve are mounted on the automatic actuator such that the fluid transfer member, the peristaltic pump, and the valve move together during movement of the automatic actuator. At least one culture medium in which the target substance is suspended is added to the plurality of containers; Multiple magnetic beads are added to the multiple containers, and the target substance is temporarily bound to the multiple magnetic beads; The plurality of containers are arranged 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 the inner surface of the respective containers within the plurality of containers; and At least the first magnetic field generating element among a plurality of magnetic field generating elements is moved in the horizontal direction relative to the working surface.

15. The purification method of claim 14, comprising causing the automanipulator to insert the end of the fluid transfer member into at least one culture medium contained in at least a first container among the plurality of containers, and removing the at least one culture medium from the first container via the fluid transfer member.

16. The purification method of claim 15, comprising, after removing the at least one culture medium from the first container, moving at least one of the plurality of magnetic field generating elements from a first position adjacent to the first container to a second position away from the first container.

17. The purification method of claim 16, comprising adding an eluent to the first container via the fluid transfer member to elute the target substance bound to the plurality of magnetic beads.

18. The purification method of claim 17, 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 the inner surface of the first container, and then removing the eluent from the first container via the fluid transfer member.

19. The purification method according to any one of claims 16 to 18, comprising adding a washing fluid to the first container via the fluid transfer member.

20. The purification method of claim 19, 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 the inner surface of the first container, and removing the washing fluid from the first container via the fluid transfer member.

21. The purification method of claim 14, wherein the plurality of magnetic field generating elements includes a first permanent magnet.

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

23. The purification method of claim 14, wherein the automated manipulator comprises a Cartesian coordinate robot capable of movement in at least the x and y directions.

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