Purification systems and methods

Abstract

A purification system and method can include a container receiving portion, a pump, and a magnetic field generating element. The container receiving portion can be configured to receive and support a container containing a mixture. Magnetic beads can be added to the container for separating a target substance from the rest of the mixture. The magnetic field generating element can be movable relative to the container receiving portion between a non-working position away from the container receiving portion and a working position adjacent an outer periphery of the container receiving portion. In the working position, the magnetic field generating element can attract the magnetic beads and hold them firmly against an inner surface of the container. The pump can remove the mixture from the container while the magnetic beads are fixed by the magnetic field generating element, leaving the magnetic beads tightly but reversibly bound to the target substance.

Description

[0001] This application is a divisional application of the original application with application number 201780062144.1.

[0002] Cross-references to related applications

[0003] The prior art claims priority to U.S. Provisional Patent Application No. 62 / 415,448, filed October 31, 2016, the entire contents of which are expressly incorporated herein by reference. Technical Field

[0004] This disclosure generally relates to purification systems and methods, and more specifically to the separation of target substances (e.g., biomolecules) from sample mixtures to facilitate analysis or further processing of the target substances. Background Technology

[0005] Many different diagnostic and medical research activities require the isolation and purification of substances (e.g., proteins contained in cell cultures or other biological mixtures). Some routine purification processes involve precipitating a target substance from a mixture by altering its solubility. Other routine purification processes require centrifugation, in which particles of different densities are separated by rotating them at high speed around a fixed point. Chromatography is another routine technique and requires passing a mixture through a medium in which its components move at different rates.

[0006] Conventional purification processes tend to be time-consuming and / or limited to relatively small sample sizes. Centrifugation is typically performed, for example, in test tubes, each of which may have a volume of less than 20 milliliters (mL). If purification of a large volume sample is required, it must usually be aliquoted into smaller sample sizes for processing using conventional purification techniques. The step of aliquoting the original sample into smaller volumes increases processing time and / or labor. Furthermore, spreading the original sample across multiple containers or wells can increase the footprint or workstation space required for the purification system. Moreover, if, for example, they require laboratory technicians to manually pipette samples between multiple different containers, then conventional purification processes can be labor-intensive.

[0007] This disclosure describes a purification system and method that provides an advantageous alternative to existing purification systems and methods, addresses one or more of the challenges or needs mentioned herein, and offers additional benefits and advantages. Summary of the Invention

[0008] One aspect of this disclosure provides a purification system that may include a container receiving portion, a first pump, and a first magnetic field generating element. The container receiving portion may be configured to receive and support a first container containing a mixture. The first pump may be configured to remove the mixture from the first container after the target substance has been separated from the remaining substances in the mixture. The first magnetic field generating element may be movable relative to the container receiving portion between a first position remote from the container receiving portion and a second position adjacent to the outer periphery of the container receiving portion.

[0009] Another aspect of this disclosure provides a purification method that may include: (a) adding a mixture comprising a target substance to a container; (b) adding magnetic beads to the container; (c) separating the target substance from the remainder of the mixture through the magnetic beads, wherein the target substance is temporarily bound to the magnetic beads; (d) applying a magnetic field to keep the magnetic beads abutting against the inner surface of the container; and (e) removing the mixture from the container while the magnetic field keeps the magnetic beads abutting against the inner surface of the container.

[0010] An additional aspect of this disclosure provides a purification system that may include a first container, a second container having a smaller volume than the first container, a first magnetic field generating element, and a second pump. The first container may be configured to initially contain a mixture and magnetic beads for separating a target substance from the remainder of the mixture. The first magnetic field generating element may be positioned adjacent to the outer surface of the second container. The first pump may be configured to transfer the mixture and the magnetic beads from the first container to the second container. The second pump may be configured to remove the mixture from the second container. Additionally, the first magnetic field generating element may magnetically attract the magnetic beads to hold them against the inner surface of the second container while the second pump removes the mixture from the second container.

[0011] Another aspect of this disclosure provides a purification method that may include: (a) adding a mixture comprising a target substance to a first container; (b) adding magnetic beads to the first container; (c) separating the target substance from the remainder of the mixture in the first container by means of the magnetic beads, wherein the target substance is temporarily bound to the magnetic beads; (d) transferring a first volume of the mixture and a first number of magnetic beads from the first container to a second container, wherein the volume of the second container is smaller than the volume of the first container; (e) applying a magnetic field to hold the first number of magnetic beads against the inner surface of the second container; (f) removing the first volume of the mixture from the second container while the magnetic field holds the first number of magnetic beads against the inner surface of the second container; (g) transferring a second volume of the mixture and a second number of magnetic beads from the first container to the second container; and (h) removing the second volume of the mixture from the second container while the magnetic field holds the first and second numbers of magnetic beads against the inner surface of the second container. Attached Figure Description

[0012] This disclosure will hopefully be more fully understood from the following description taken in conjunction with the accompanying drawings. Some drawings may have been simplified by omitting selected elements for the purpose of illustrating other elements more clearly. Such omission of elements in some drawings does not necessarily indicate the presence or absence of a particular element in any embodiment, unless it can be explicitly described in the corresponding written description. Moreover, these drawings are not necessarily drawn to scale.

[0013] Figure 1 This is a schematic top view representing an embodiment of a purification system constructed according to the principles of this disclosure.

[0014] Figure 2 This is a side perspective view of an embodiment of a purification system constructed according to the principles of this disclosure, wherein the magnetic field generating elements are arranged in their respective non-operating positions.

[0015] Figure 3 yes Figure 2 The image depicts a side perspective view of the purification system, in which the magnetic field generating elements are arranged in their respective working positions.

[0016] Figures 4A to 4H An embodiment of a purification method based on the principles of this disclosure is illustrated schematically.

[0017] Figure 5 This is a schematic top view representing another embodiment of a purification system constructed according to the principles of this disclosure.

[0018] Figure 6A and Figure 6B Another embodiment of the purification method based on the principles of this disclosure is illustrated schematically. Detailed Implementation

[0019] Figure 1 This is a schematic representation of one embodiment of the purification system 10, which, according to the principles of this disclosure, can be used to separate a target substance from a mixture. The purification system 10 may include a container receiving portion 12 configured to receive and support a container 14, a pump 16 in fluid communication with the container 14 via a fluid conduit 18, a plurality of magnetic field generating elements 20a-d, and a plurality of actuators 22a-d, each configured to move a corresponding magnetic field generating element among the plurality of magnetic field generating elements 20a-d relative to the container receiving portion 12. The container 14 may be filled with a sample mixture 30 including the target substance T. A plurality of magnetic beads 32 may be submerged (e.g., suspended) in the sample mixture 30 to separate or isolate the target substance T from the remaining substances in the sample mixture 30. Additionally, the pump 16 may be fluidly connected to a multi-position valve 24, allowing the pump 16 to selectively discharge waste to a drain 26 and deliver rinsing fluid, eluent, and / or other fluids from one or more additional containers 28a and 28b to the container 14. In addition, the purification system 10 may include a control unit 29 configured to automatically control the operation of the pump 16 and / or actuators 22a-d.

[0020] Generally, during the operation of the purification system 10, container 14 can be placed in container receiving section 12, and depending on the stage of the purification process, actuators 22a-d can cause each of the magnetic field generating elements 20a-d to reciprocate between a non-operating position away from container 14 and an adjacent (e.g., adjacent but spaced apart, or adjacent and in direct contact) operating position of container 14 and / or container receiving section 12. When the magnetic field generating elements 20a-20d are arranged in their respective adjacent operating positions of container 14, they can attract magnetic beads 32 and keep them abutting against the inner surface 34 of container 14. This allows pump 16 to remove sample mixture 30 from container 14, while the magnetic beads 32 bound to the target substance T remain in container 14 for subsequent rinsing and / or elution procedures.

[0021] With this configuration, the purification system and basic method disclosed herein advantageously provide an automated or semi-automated process for separating target substances from mixtures. Furthermore, by virtue of the ability to apply a relatively strong magnetic field and / or surround the container with multiple magnetic field generating elements, the purification system disclosed herein can process sample mixtures with relatively large volumes and containing numerous magnetic beads, including samples with volumes, for example, equal to or greater than 1 liter (L). Accordingly, the purification system and method disclosed herein can eliminate or reduce the need to aliquot large volumes of samples into multiple processing containers (as done in many conventional purification systems and methods). Consequently, the purification system and method disclosed herein can advantageously provide a faster or more efficient method for purifying mixtures, and can also reduce the floor space or workstation space required to house the purification equipment. Additionally, as will be further described below, the purification system disclosed herein can be configured for continuous flow applications, in which the container and magnetic field generating elements generally function as traps for removing magnetic beads from a continuous stream of sample mixtures containing magnetic beads. This allows for the processing of very large samples (e.g., samples with a volume exceeding 20,000 L) while minimizing the amount of rinsing solution and / or eluent required to extract the target material bound to these magnetic beads.

[0022] Each of the aforementioned components of the purification system 10 and the associated purification methods will now be described in more detail.

[0023] The purification system and method disclosed herein can be used to separate 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, saliva, mucus, sweat or perspiration, urine, feces, soil, food products, etc.). Moreover, a wide variety of magnetic beads can be used depending on the nature of the target substance T to be separated and / or the sample mixture 30. In some embodiments, these magnetic beads 32 may have a spherical shape and a paramagnetic core encapsulated in silica, and the beads may be coated with a material that binds to or is coupled to the target substance T. The binding action between the magnetic beads 32 and the target substance T can be achieved covalently, non-covalently, or electrostatically via hydrogen bonding, van der Waals forces, and / or any other suitable molecular binding method. In at least one embodiment, the sample mixture 30 may be a cell culture, the target substance T in the sample mixture 30 may be an antibody, and the magnetic beads 32 may be protein A beads. In another embodiment, the sample mixture 30 may be a cell culture, the target substance T in the sample mixture 30 may be an HIS-labeled protein, and the magnetic beads 32 may be coated with zinc, copper, or cobalt for protein purification.

[0024] 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, acetaldehyde magnetic beads, carboxyl magnetic beads, CDI magnetic beads, DVS magnetic beads, DADPA magnetic beads, epoxy magnetic beads, hydrazide magnetic beads, hydroxy magnetic beads, iodoacetyl magnetic beads, NHS magnetic beads, thiol 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 type magnetic beads (e.g., protein A magnetic beads, protein G magnetic beads, protein A / G magnetic beads, protein L magnetic beads, Quick IgG Pure Magnetic beads). Beads, antigen peptide magnetic beads, Quick IgM Pure Magnetic Beads, Anti-IgG Magnetic Beads, Quick IgA Pure Magnetic Beads Magnetic beads for DNA or RNA purification (e.g., protein A beads, protein G beads, protein A / G beads, protein L beads, epoxy activated beads, acetaldehyde-terminated beads, hydrazide-terminated beads, carboxyl-terminated beads, iodoacetyl-activated beads, thiol-activated beads, etc.); magnetic beads for recombinant protein purification (e.g., Ni+-banded magnetic beads, Co+-banded magnetic beads, maltose beads, calmodulin beads, etc.); magnetic beads for peptide immobilization (e.g., epoxy activated beads, acetaldehyde-terminated beads, carboxyl-terminated beads, amino-terminated beads, iodoacetyl-activated beads, thiol-activated beads, etc.); magnetic beads for DNA or RNA purification; magnetic beads for endotoxin removal; magnetic beads for protein enrichment and removal; and / or EDTA beads.

[0025] As used herein, the term "magnetic" is defined to encompass any element that is magnetic, paramagnetic, and / or ferromagnetic. Accordingly, the magnetic bead 32 may be a magnetic bead, a paramagnetic bead, a ferromagnetic bead, or any combination thereof.

[0026] In some embodiments, the magnetic beads 32 may have a density greater than that of the sample mixture 30, such that when the magnetic beads 32 are submerged in the sample mixture 30, the magnetic beads 32 sink to the bottom of the container 14. In other embodiments, the magnetic beads 32 may have a density less than or equal to that of the mixture 30, such that the magnetic beads 32 float or partially float in the sample mixture 30.

[0027] Still refer to Figure 1The container receiving portion 12 may include a platform 41 for receiving and supporting the container 14. In some embodiments, the container receiving portion 12 may be equipped with an engagement structure 42 for suppressing lateral movement of the container 14 when it is arranged on the platform 41. Figure 2 and Figure 3 (Not shown in the image). In some embodiments, the engagement structure 42 may include one or more vertical walls arranged around the perimeter of the platform 41 and forming a sleeve around the container 14. Each of these vertical walls may have a height approximately equal to the height of the container 14. Alternatively, the engagement structure 42 may be a relatively short, vertical edge surrounding the base of the container 14. The engagement structure 42 may be useful in suppressing movement of the container 14 when the magnetic beads 32 initially move into contact with the inner surface of the container 14 due to the magnetic attraction of the magnetic field generating elements 20a-d. In embodiments where the container receiving portion 12 includes the engagement structure 42, the outer perimeter or boundary of the container receiving portion 12 may be defined by the engagement structure 42. If the engagement structure 42 is omitted, the outer edge of the platform 41 itself may define the outer perimeter or boundary of the container receiving portion 12. Furthermore, in some embodiments, the engagement structure 42 may act as a stop to prevent the magnetic field generating elements 20a-d from advancing further into the container 14.

[0028] Reference Figure 2 The diagram shows a perspective view of one embodiment of the purification system 10, wherein each of the magnetic field generating elements 20a-d is arranged in a non-operating position away from the container 14. The purification system 10 may be incorporated with a frame 40 or housing to connect and / or support multiple different components, including the container receiving portion 12, the pump 16, the magnetic field generating elements 20a-d, and the actuators 22a-d. In this embodiment, the pump 16 is mounted on top of the frame 40 such that it is arranged vertically above the container receiving portion 12. The frame 40 may include channels or slots 43a-d that define a fixed horizontal path for each of the magnetic field generating elements 20a-d. These paths are straight and each aligned with an imaginary radial line emanating from the center C of the container receiving portion 12. However, in other embodiments, the paths defined by the slots 43a-d may be non-linear, curved, or have any other suitable shape. Furthermore, in some embodiments, the frame 40 may be incorporated with an outer shell structure (not shown) for creating a sealable internal space in which other components of the purification system 10 can be housed. The outer shell structure may include a door for accessing the internal space, for example, for adding and removing container 14.

[0029] Still refer to Figure 2The container 14 may include a bottom wall 50 defining a closed end of the container 14, a side wall 52 extending upward from the bottom wall 50 and defining an internal volume V1 of the container 14, and a top wall 54 having an opening 56 defining an open end of the container 14. The side wall 52 may define an inner surface 34 and an outer surface 57 of the container 14. A longitudinal axis A of the container 14 may extend between the bottom wall 50 or the closed end of the container 14 and the top wall 54 or the open end of the container 14. In some embodiments, the top wall 54 may be funnel-shaped to form a pouring outlet. Figure 1 In the illustrated embodiment, the top wall 54 can be omitted, allowing the upper end of the container 14 to be entirely defined by the opening 56. A removable cap 58 can cover the opening 56 and can be removed during the purification process, thereby allowing the fluid conduit 18 ( Figure 2 (Not shown) can be inserted into container 14. Container 14 can be made of any suitable material, including, for example, glass or plastic. Figure 2 In the illustrated embodiment, container 14 is made of a transparent material. Furthermore, in some embodiments, container 14 may be a large laboratory beaker.

[0030] In this embodiment, container 14 has a generally cylindrical shape with a diameter D. The diameter D can range from approximately (e.g., ±10%) 10-60 cm, or approximately (e.g., ±10%) 20-50 cm, or approximately (e.g., ±10%) 35-45 cm, or greater than or equal to approximately (e.g., ±10%) 10 cm, or greater than or equal to approximately (e.g., ±10%) 20 cm, or greater than or equal to approximately (e.g., ±10%) 30 cm. In other embodiments, container 14 may have a square or rectangular cross-sectional shape, or any other suitable cross-sectional shape. While container 14 in this embodiment is configured to keep mixture 30 in a non-flowing state when magnetic beads 32 are removed, in alternative embodiments, container 14 may be configured to allow continuous or semi-continuous flow of mixture 30 and / or other fluids through a conduit (e.g., a tube) when magnetic beads 32 are removed and / or captured.

[0031] Generally, the volume V1 of container 14 can be larger than the volume of containers or sample wells used in conventional purification systems. In some embodiments, the volume V1 of container 14 can range from about (e.g., ±10%) 0.5-5.0 L, or about (e.g., ±10%) 0.5-4.0 L, or about (e.g., ±10%) 0.5-3.0 L, or about (e.g., ±10%) 0.5-2.0 L, or about (e.g., ±10%) 1.0-3.0 L, or about (e.g., ±10%) 1.0-2.0 L, or greater than or equal to about (e.g., ±10%) 0.5 L, or greater than or equal to about (e.g., ±10%) 1.0 L. This allows for the purification of sample mixtures with relatively large volumes without distributing the sample mixture 30 among multiple processing containers or sample wells. Alternatively, the purification process can occur in a single container (i.e., container 14), and in some cases, the same container that previously stored the sample mixture can be used during the purification process. Due to the relatively large volume of container 14, purification system 10 is able to purify the sample mixture up to 10 times faster than conventional purification systems, or even faster.

[0032] like Figure 1 As shown, pump 16 can be in fluid communication with container 14 via fluid conduit 18. Fluid conduit 18 can be made of a flexible tube such that when container 14 is placed on container receiving portion 12, fluid conduit 18 can bend laterally and then backward to its initial position with the distal end of fluid conduit 18 inserted into the interior of container 14. Figure 2 As shown, pump 16 can be mounted on frame 40, such that pump 16 is vertically positioned above container 14. In other embodiments, pump 16 may be laterally positioned to the side of container 14 and / or not connected to frame 40.

[0033] Generally, pump 16 is configured to remove fluid via fluid conduit 18 and / or add fluid to container 14. Pump 16 can be powered by any suitable device, including but not limited to electric motors and / or pressurized hydraulic fluid and / or gas sources. Depending on the specifications of the purification process, pump 16 can be operated at a variable speed or a single speed. In some embodiments, the operation of pump 16 can be electronically controlled by control unit 29 according to programmable instructions, for example, stored in the memory of control unit 29. Alternatively, or additionally, pump 16 can be operated by an operator (e.g., a laboratory technician) who manually actuates ON / OFF switch 46 and / or rotates speed knob 48, such as... Figure 2As illustrated in the embodiments. Furthermore, in some embodiments, pump 16 may be a positive displacement pump (e.g., a peristaltic pump) and capable of pumping fluids containing suspended solids (e.g., magnetic beads) without causing damage. The continuous flow type purification process described below can utilize this embodiment of pump 16, where the mixture and magnetic beads are transferred together from a culture vessel or tank to container 14. In yet another embodiment, pump 16 may be a centrifugal pump (e.g., a radial flow pump) that uses a rotating impeller to create a vacuum to move the fluid. Additionally, in some embodiments, pump 16 may be reversible.

[0034] like Figure 2 and Figure 3 As shown, the multi-position valve 24 can be built into the housing containing the pump 16, or alternatively, the multi-position valve 24 can be as follows: Figure 1 As shown, it is separate from pump 16. Multi-position valve 24 may allow pump 16 to be selectively fluidly connected to various different containers (e.g., containers 28a and 28b) and / or drain 26. Figure 1 In the illustrated embodiment, the multi-position valve 24 is a 3-way valve configured to selectively connect pump 16 to drain 26, container 28a, or container 28b. In other embodiments, the multi-position valve 24 may be a 2-way valve, or any other valve having any number of selectively openable orifices. In some embodiments, the operation of the multi-position valve 24 may be electronically controlled by control unit 29 according to programmable instructions, for example, stored in the memory of control unit 29. The multi-position valve 24 may include one or more solenoids for opening and closing the orifices of the multi-position valve 24 in response to command signals from control unit 29. In other embodiments, the multi-position valve 24 may be manually controlled by a user, for example, by rotating a rotary valve. Figure 2 and Figure 3 The valve knob 48 protrudes from the pump housing as shown. In an alternative embodiment, the second multi-position valve may include a valve configured to selectively connect pump 16, or the second pump fluid, to one of several drains or other destinations for fluid drawn from container 14.

[0035] Reference Figure 2 The purification system 10 may include a rack 50 for holding a plurality of different containers, including containers 28a and 28b. In some embodiments, one of the containers held by the rack may be used as... Figure 1 The drain element 26 is shown. The rack 50 can be rigidly connected to or detached from the frame 40. The container held by the rack 50 can contain rinsing solutions, eluents, buffers, enzymes, and / or other reagents to be used during the purification process. Furthermore, the rack 50 can be configured to hold the sample mixture container 14 before the purification process begins.

[0036] Figure 2 and Figure 3 The diagram shows magnetic field generating elements 20a-d arranged at equal intervals around the outer perimeter of the container receiving portion 12. In the illustrated embodiment, four magnetic field generating elements 20a-d are included, such that the interval between each of these magnetic field generating elements 20a-d is 90 degrees. In other embodiments, two, three, five, six, seven, eight, nine, ten, or more magnetic field generating elements may be included. By using multiple magnetic field generating elements around the container 14, a more uniform magnetic field can be generated when the magnetic field generating elements are arranged in their respective operating positions (see...). Figure 3 In this configuration, the magnetic field helps ensure that all the magnetic beads 32 remain and are fixed against the inner surface 34 of the container 14. In an embodiment that includes only two magnetic field generating elements, each magnetic field generating element may have a C-shaped cross-section, such that moving the two magnetic field generating elements to their respective working positions causes them to surround the container 14. In a further embodiment, only a single magnetic field generating element may be included, and although it is located on only a single side of the container 14, it can generate a sufficiently strong magnetic field to magnetically attract most or all of the magnetic beads 32 against the inner surface 34 of the container 14.

[0037] In some embodiments (such as one embodiment shown in the accompanying drawings), each of these magnetic field generating elements 20a-d is composed of a corresponding permanent magnet configured to generate its own persistent magnetic field. Each of these permanent magnets may have a maximum magnetic pull force ranging from approximately (e.g., ±10%) 50-1000 Newtons (N), or approximately (e.g., ±10%) 100-800 N, or approximately (e.g., ±10%) 100-700 N, or approximately (e.g., ±10%) 150-600 N, or approximately (e.g., ±10%) 200-500 N, or approximately (e.g., ±10%) 200-450 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 approximately (e.g., ±10%) 500 N, or greater than or equal to approximately (e.g., ±10%) 1000 N, or greater than or equal to approximately (e.g., ±10%) 1500 N, or greater than or equal to approximately (e.g., ±10%) 2000 N, or greater than or equal to approximately (e.g., ±10%) 2500 N. In some embodiments, the permanent magnets constituting the magnetic field generating elements 20a-d may be nickel-plated neodymium magnets, each having a rectangular shape generally approximately (e.g., ±10%) 5 cm long, 5 cm wide, and 1.25 cm thick. In alternative embodiments, each of these magnetic field generating elements 20a-d may be composed of a corresponding electromagnet configured to generate a magnetic field when supplied with current. In such embodiments, the power supply to the electromagnet may be automatically controlled by the control unit 29.

[0038] Each of these magnetic field generating elements 20a-d can be located relative to the container receiving portion 12 and / or the container 14 from a corresponding non-operating position remote from the container receiving portion 12 and / or the container 14 (see...). Figure 2 Move to the corresponding adjacent container receiving section 12 and / or container 14 (see Figure 3The working position of the magnetic field generating elements 20a-d is located at the outer periphery of the container 14. Movement between the working and non-working positions can generally be in the horizontal direction and / or in a direction not parallel (e.g., perpendicular) to the longitudinal axis A of the container 14. The advantage of laterally moving the magnetic field generating elements 20a-d into and out of the container 14 is that it does not obstruct the opening 56 in the top of the container 14 and also eliminates the need to submerge the magnetic field generating elements 20a-d in the sample mixture 30 to attract the magnetic beads 32 (which carries the risk of introducing contaminants). In the non-working position, the magnetic field of each of the magnetic field generating elements 20a-d near the container 14 may be too weak to affect the position of the magnetic beads 32 within the container 14. In the working position, the magnetic field of the magnetic field generating elements 20a-d near the container 14 can be strong enough to magnetically attract the magnetic beads 32 and hold them against the inner surface 34 of the container 14, thereby fixing the magnetic beads 32 relative to the container 14. In some embodiments, the distance between a non-working position and its corresponding working position may fall within the following ranges: approximately (e.g., ±10%) 10-75 cm, or approximately (e.g., ±10%) 20-60 cm, or approximately (e.g., ±10%) 30-45 cm, or greater than or equal to approximately (e.g., ±10%) 20 cm, or greater than or equal to approximately (e.g., ±10%) 30 cm.

[0039] like Figures 1 to 3 As shown, each of these magnetic field generating elements 20a-d can be configured to move along a fixed path of a corresponding straight line, which is aligned with a corresponding imaginary radial line emanating from the center C of the container receiving portion 12 and / or the longitudinal axis A of the container 14 (which may pass through the center C of the container receiving portion 12). In a general sense, the fixed paths of the straight lines of the magnetic field generating elements 20a-d can be arranged like spokes on a wheel. Accordingly, each of these magnetic field generating elements 20a-d can move inward toward the center C of the container receiving portion 12 when moving from its non-working position to its working position, and can move outward away from the center C of the container receiving portion 12 when moving from its working position to its non-working position. Therefore, the distance between the container 14 and each of the magnetic field generating elements 20a-d in its non-working position can be greater than the distance between the container 14 and each of the magnetic field generating elements 20a-d in its working position. In some embodiments, the fixed paths of these magnetic field generating elements 20a-d may be non-linear, curved, or have any other suitable shape.

[0040] Furthermore, each of these magnetic field generating elements 20a-d may have a corresponding actuator 22a-d configured to move the magnetic field generating element back and forth between a non-operating position and an operating position. In some embodiments, each of these actuators 22a-d may include a hydraulic cylinder or a pneumatic cylinder with a reciprocating piston rod. In such embodiments, a single source (not shown) of pressurized hydraulic fluid or gas can power each of these hydraulic or pneumatic cylinders. In other embodiments, each of these actuators 22a-d may be powered by a rotary electric motor (not shown), for example, a pulley system, a gear system, or some other motion conversion mechanism. Including actuators 22a-d capable of automatically moving the magnetic field generating elements 20a-d can be advantageous because operating the magnetic field generating elements 20a-d may pose a danger to the operator's health and / or the memory of personal electronic devices carried by the operator. Furthermore, if the magnetic attraction between the magnetic bead 32 and the magnetic field generating elements 20a-d is very strong, or if the magnetic attraction between these magnetic field generating elements 20a-d themselves is very strong, then manually moving the magnetic field generating elements 20a-d from the working position to the non-working position may be difficult and / or cumbersome for the operator. However, in some embodiments, it is possible that the actuators 22a-d may not be automatic, and instead may be manually powered by the user via, for example, a crank mechanism, or simply by the user pushing and pulling the actuators 22a-d. In some embodiments, the actuators 22a-d may be configured to move the magnetic field generating elements 20a-d uniformly; while in other embodiments, the actuators 22a-d may be configured to move the magnetic field generating elements 20a-d independently of each other.

[0041] like Figure 1As shown, the purification system 10 may include a control unit 29 for controlling the operation of the pump 16, actuators 22a-d, and / or other controllable elements of the purification system 10. The control unit 29 may include a processor (e.g., a microprocessor), memory for storing tangible, non-transitory computer-readable instructions (e.g., random access memory (RAM), non-persistent 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, buttons, a touchscreen, etc.). Furthermore, in some embodiments, the control unit 29 may be a programmable logic controller. The control unit 29 may be programmed to perform the purification process according to predetermined specifications of the operator. In some embodiments, the control unit 29 may perform steps of the purification process, such as activating actuators 22a-d to move magnets 20a-d in response to properties of the sample mixture 30 represented by sensor data (e.g., volume, temperature, weight, pH, etc.), timers, analog or digital input from the operator, and / or any other relevant detectable item or event.

[0042] Now refer to Figures 4A-4H The operating method of purification system 10 is described. For discussion purposes, Figures 4A to 4H Some components of purification system 10 are not depicted in the figures. Their omission in the figures should not be construed as meaning they are necessarily absent from purification system 10. In the initial step, container 14 may be filled with a volume of sample mixture 30 containing the target substance T. Next, as... Figure 4A As shown, magnetic beads 32 can be added to container 14 and left to interact with sample mixture 30 for a period of time (e.g., minutes, hours, days, etc.). During this incubation period, magnetic beads 32 can bind to target substance T, thereby separating target substance T from the rest of the sample mixture 30 (see...). Figure 4B As previously described, the binding of magnetic beads 32 to 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 method. In some embodiments, the sample mixture 30 can be agitated or shaken during the incubation period to promote the binding of the target substance T to the magnetic beads 32. Furthermore, in some embodiments, when the magnetic beads 32 are bound to the target substance T, the container 14 can be placed on the container receiving portion 12.

[0043] Next, if this has not already been done, container 14 can be placed on container receiving portion 12, while magnetic field generating elements 20a-d are each arranged in their respective non-operating positions, such as... Figure 4CAs shown. In some embodiments, container 14 may be secured to container receiving portion 12 by a removable double-sided adhesive pad. During this step, the distal end of fluid conduit 18 may be inserted into container 14 through opening 56 and submerged in sample mixture 30. The opening of fluid conduit 18 may be positioned adjacent to or otherwise very close to the bottom wall 50 of container 14, such that fluid conduit 18 can aspirate all or substantially all of sample mixture 30 from container 14. In alternative embodiments, fluid conduit 18 may be fluidly connected to an opening (not shown) in the bottom wall 50 or side wall 52 of container 14.

[0044] Subsequently, as Figure 4D As shown, actuators 22a-d can be activated to move magnetic field generating elements 20a-d inward from their respective non-operating positions away from container 14 to their respective operating positions adjacent to (e.g., adjacent but spaced apart, or adjacent and in direct contact with) the sidewall 52 of container 14 and / or the outer periphery of container receiving portion 12. Although Figure 4D Only magnetic field generating elements 20a and 20b are depicted; however, at this stage, all magnetic field generating elements 20a-d can be moved from their respective non-operating positions to their respective operating positions. As a result, magnetic field generating elements 20a-d can magnetically attract the magnetic bead 32, thereby causing the magnetic bead 32 to remain against the inner surface 34 of the sidewall 52 of the container 14. The friction or contact between the magnetic bead 32 and the magnetized inner surface 34 of the sidewall 52 of the container 14 can effectively fix or bind the magnetic bead 32 relative to the container 14, thereby suppressing or preventing movement or dispersion of the magnetic bead 32 during subsequent fluid removal or addition to the container 14. In some embodiments, actuators 22a-d can automatically move the magnetic field generating elements 20a-d in response to a command signal from the control unit 29. In other embodiments, actuators 22a-d can be manually powered by an operator to move the magnetic field generating elements 20a-d.

[0045] Reference Figure 4E The next step in the purification process may involve aspirating or removing the sample mixture 30 from the container 14 via the fluid conduit 18. In some embodiments, the pump 16 may be used to generate the suction required to remove the sample mixture 30 from the container 14. The magnetic beads 32 are not removed in this step because they are held against the inner surface 34 of the sidewall 52 of the container 14 by the magnetic field generating elements 20a-20d.

[0046] After removing sample mixture 30, as Figure 4FAs shown, rinsing fluid 60 (e.g., a salt solution) can be added to container 14 via fluid conduit 18 or another fluid conduit to rinse the inner surface 34 of container 14 and / or any remaining mixture of magnetic beads 32. However, rinsing fluid 60 may not remove the target substance T from the magnetic beads 32. In some embodiments, rinsing fluid 60 can be pumped from one of the containers (e.g., containers 28a or 28b) held by rack 50 to container 14 by pump 16. The volume of rinsing fluid 60 added to container 14 can be equal to or less than the maximum volume V1 of container 14 (e.g., a portion thereof). Optionally, during the rinsing step, actuators 22a-d can return magnets 20a-d to their non-operating positions, thereby allowing the magnetic beads 32 to be freely dispersed (e.g., suspended) in the rinsing fluid 60. Allowing the magnetic beads 32 to disperse can increase the effectiveness of the rinsing process and / or allow stirring, mixing, or shaking of the magnetic beads 32 in the rinsing fluid 60. Once rinsing is complete, actuators 22a-d can move magnets 20a-d back to their working positions to once again secure the magnetic bead 32 against the inner surface 34 of the container 14, and the rinsing fluid 60 can be removed. Furthermore, in some embodiments, the rinsing step described above can be omitted.

[0047] Next, as Figure 4G As shown, eluent 62 (e.g., a fluid elution solution) can be added to container 14 via fluid conduit 18 or another fluid conduit to release the target substance T from the magnetic beads 32. Actuators 22a-d can be configured to move magnetic field generating elements 20a-d to their respective non-operating positions during the elution step, thereby allowing the magnetic beads 32 to be freely dispersed (e.g., suspended) in the eluent 62; however, it is not required to move the magnetic field generating elements 20a-d to their non-operating positions at this stage. In some embodiments, eluent 62 can be transferred from one of the containers (e.g., container 28a or 28b) held by rack 50 to container 14 by pump 16. In some embodiments, the volume of eluent 62 transferred to container 14 by pump 16 can be equal to or less than the volume of rinsing solution 60 previously added to container 14 during the rinsing step. Furthermore, in some embodiments, an additional step of adding enzyme and / or buffer components may be included to, for example, expand the target substance T in the eluent 62.

[0048] After the incubation period, such as Figure 4HAs shown, actuators 22a-d can move magnets 20a-d back to their respective working positions to re-secure magnetic beads 32 against the inner surface 34 of container 14. Pump 16 can then remove eluent 62 and the target substance T bound to eluent 62 from container 14 via fluid conduit 18 and transfer them to one container or the other held by rack 50. Finally, container 14 can be removed from container receiving section 12 and discarded or stored for future use. The above steps can then be repeated for another container containing a different mixture requiring purification.

[0049] Although this embodiment uses pump 16 to add fluid and remove fluid from container 14, in alternative embodiments, an operator may manually perform either the fluid addition or removal step using, for example, a pipette.

[0050] Figure 5 This is a schematic representation of another embodiment of the purification system 110 constructed according to the principles of this disclosure. The purification system 110 is similar in some respects to the purification system 10 described above, except that a culture vessel or tank 180 and a transfer pump 182 are added to facilitate continuous flow processing of large volumes of sample mixtures. Components of the purification system 110 that are the same as or similar to the purification system 10 are indicated by the same reference numerals plus 100. For the sake of brevity, descriptions of many of these components have been omitted or omitted.

[0051] Overall, the incubation container 180 provides holding space for the large volume sample mixture 130, while the magnetic beads 132 submerged in the sample mixture 130 separate the target substance T from the rest of the sample mixture 130. The volume V2 of the incubation container 180 can be significantly larger than the volume V1 of the container 114. In some embodiments, the volume V2 of the incubation container 180 can be at least 5 times, at least 10 times, at least 100 times, or even more than a multiple of the volume V1 of the container 114.

[0052] The transfer pump 182 can be configured to transfer the mixture 130 and the magnetic beads 132 from the culture container 180 to the container 114, which in this embodiment generally serves as a trap for removing the magnetic beads 132 from the mixture 130. The transfer pump 182 can be fluidly connected to the culture container 180 via a fluid conduit 184 (e.g., a flexible tube) and to the container 114 via a fluid conduit 186 (e.g., a flexible tube). In some embodiments, the transfer pump 182 can be mounted on a frame 150.

[0053] Transfer pump 182 can be configured to remove fluid via fluid conduit 184 and / or add fluid to culture vessel 180. Transfer pump 182 can be powered by any suitable device, including but not limited to electric motors and / or pressurized hydraulic fluid and / or gas sources. Depending on the specifications of the purification process, transfer pump 182 can be operated at a variable speed or a single speed. In some embodiments, operation of transfer pump 182 can be electronically controlled by control unit 29 according to programmable instructions, for example, stored in the memory of control unit 29. Alternatively, or additionally, transfer pump 182 can be operated manually, for example, by an operator (e.g., a laboratory technician) actuating an ON / OFF switch and / or rotating a speed knob. Furthermore, in some embodiments, transfer pump 182 can be a positive displacement pump (e.g., a peristaltic pump) and is capable of pumping magnetic beads 132 suspended in mixture 130 without damaging these beads 132. In yet another embodiment, transfer pump 182 can be a centrifugal pump (e.g., a radial flow pump) that uses a rotating impeller to create a vacuum to move the fluid. In addition, in some embodiments, the transfer pump 182 may be reversible.

[0054] exist Figure 5 In the embodiments shown, similar to the above combination Figures 1 to 4H The described embodiments include actuators 122a-d to move magnetic field generating elements 120a-d back and forth between an operating position and a non-operating position. However, in other embodiments, actuators 122a-d may be omitted, and magnetic field generating elements 120a-d may be permanently or otherwise fixedly positioned at their respective operating positions adjacent to (e.g., adjacent but spaced apart, or adjacent and in direct contact with) the outer surface 157 of the sidewall 152 of the container 114.

[0055] Now refer to Figure 6A and Figure 6B Describe the operating procedure of purification system 110. For discussion purposes, Figure 6A and Figure 6B Some components of purification system 110 are not depicted in the figures. Their omission in the figures should not be interpreted as meaning that they are necessarily missing from purification system 110.

[0056] Initially, sample mixture 130 can be added to incubation container 180, followed by the addition of magnetic beads 132. Magnetic beads 132 can be left to interact with sample mixture 130 for a period of time (e.g., minutes, hours, days, etc.). During this incubation period, magnetic beads 132 can bind to target substance T, thereby separating target substance T from the remaining substances in sample mixture 130.

[0057] Next, the transfer pump 182 can be activated to begin pumping a first volume of sample mixture 130 and a first number of magnetic beads 132 suspended in the first volume of sample mixture 130 from the culture container 180 to the container 114, as follows. Figure 6A As shown. Simultaneously or shortly after activating transfer pump 182, pump 116 can be activated to begin removing sample mixture 130 from container 114 and discharging it, for example, into waste container or drain 126. Before activating either pump 116 or 182, magnetic field generating elements 120a-d can be positioned in their respective working positions adjacent to (e.g., adjacent but spaced apart, or adjacent and in direct contact with) the outer surface 157 of the sidewall 152 of container 114. In some embodiments, this positioning of magnetic field generating elements 120a-d can be accomplished via actuators 122a-d in a manner similar to that discussed above. In other embodiments, magnetic field generating elements 120a-d can be permanently or otherwise fixedly positioned in their respective working positions. In either case, magnetic field generating elements 120a-d can magnetically attract magnetic beads 132 added to container 114, thus causing magnetic beads 132 to be held against the inner surface 134 of the sidewall 152 of container 114. When the magnetic field generating elements 120a-d are in their respective operating positions, the friction or contact between the magnetic bead 132 and the inner surface 134 of the sidewall 152 of the container 114 can effectively fix or bind the magnetic bead 132 relative to the container 114. Accordingly, the magnetic bead 132 is firmly anchored to the wall of the container 114, and thus inhibits or prevents the magnetic bead from being removed from the container 114 during the removal of the sample mixture 130 from the container 114 by the pump 116. It should be noted that, although Figure 6A and Figure 6B Only magnetic field generating elements 120a and 120b are depicted, but all magnetic field generating elements 120a-d can be positioned in their respective operating positions at this stage of processing.

[0058] Subsequently, transfer pump 182 can pump a second volume of sample mixture 130 and a second number of magnetic beads 132 suspended in the second volume of sample mixture 130 from incubation container 180 to container 114. After pumping the first volume of sample mixture 130 and the first number of magnetic beads 132, a second volume of sample mixture 130 and the second number of magnetic beads 132 can be pumped immediately, thereby achieving uninterrupted continuous flow of sample mixture 130 and magnetic beads 132 from incubation container 180 to container 114. Furthermore, pump 116 can be operated to remove sample mixture 130 from container 114, while simultaneously, a second volume of sample mixture and the second number of magnetic beads 132 are pumped into container 114 by transfer pump 182. Accordingly, uninterrupted continuous flow of sample mixture 130 through container 114 can be achieved. Throughout this process, as... Figure 6BAs shown, the magnetic bead 132 is continuously fixed against the inner surface 134 of the side wall 152 of the container 114.

[0059] In some embodiments, transfer pump 182 and pump 116 may be configured (e.g., controlled by control unit 129) such that the volumetric flow rate of sample mixture 130 through transfer pump 182 is equal to or approximately equal to the volumetric flow rate of sample mixture 130 through transfer pump 116. Accordingly, throughout the magnetic bead removal process, except initially (when container 114 is initially filled by transfer pump 182) and finally (when container 114 is completely drained by pump 114), the fluid level in container 114 may remain relatively constant.

[0060] Once the material has been transferred from the cultivation container 180 to the capture container 114, the pump 114 can drain all remaining mixture 130 from container 114, leaving only the magnetic beads 132 bound to the target substance T in container 114. Subsequently, the above-mentioned binding process can be performed in container 114. Figure 4F , 4G The rinsing and elution steps discussed in 4H are similar to those in 4H.

[0061] The continuous flow aspect of this purification method makes it conveniently applicable to purifying very large sample mixtures, including those exceeding 20,000 L. Furthermore, since all magnetic beads 132 are simultaneously rinsed and / or eluted in a single container, the amount of rinsing and / or eluent is reduced compared to aliquoting large volume sample mixtures into numerous processing containers.

[0062] While the systems and methods disclosed herein have been described in conjunction with several different embodiments, it should be understood that the systems and methods disclosed herein can be further modified. This application is intended to cover any variations, uses, or adaptations of systems and methods that generally follow the principles of this disclosure, and includes differences from this disclosure in known and customary practices falling within the field to which this invention pertains.

Claims

1. A purification method, comprising: Add the mixture, including the target substance, to the container; Add the magnetic beads to the container; The target substance is separated from the remaining substances in the mixture by means of the magnetic beads, wherein the target substance is temporarily bound to the magnetic beads; Multiple magnetic field generating elements are arranged around the container, and these elements are simultaneously moved toward the container to apply a magnetic field, thereby keeping the magnetic bead in contact with the inner surface of the container; and The mixture is removed from the container while the magnetic field keeps the magnetic beads against the inner surface of the container.

2. The purification method of claim 1, comprising adding a rinsing solution to the container after removing the mixture from the container, and subsequently removing the rinsing solution from the container while the magnetic field keeps the magnetic beads against the inner surface of the container.

3. The purification method of claim 2, comprising removing the magnetic field after adding the rinsing solution to the container, such that the magnetic beads are freely dispersed in the rinsing solution, and subsequently reapplying the magnetic field during removal of the rinsing solution from the container to keep the magnetic beads against the inner surface of the container.

4. The purification method according to any one of claims 1 to 3, comprising adding an eluent to the container after removing the mixture from the container to elute the target substance bound to the magnetic beads, and subsequently removing the eluent and the target substance from the container while the magnetic field keeps the magnetic beads against the inner surface of the container.

5. The purification method of claim 4, further comprising removing the magnetic field after adding the eluent to the container, such that the magnetic beads are freely dispersed in the eluent, and subsequently reapplying the magnetic field to keep the magnetic beads against the inner surface of the container during removal of the eluent from the container.

6. The purification method of claim 1, wherein, Removing the mixture from the container includes pumping the mixture from the container via an automatic pump.

7. The purification method of claim 1, wherein, Moving the plurality of magnetic field generating elements simultaneously toward the container to apply the magnetic field to keep the magnetic bead abutting the inner surface of the container includes: activating an actuator to move the plurality of magnetic field generating elements from a first position away from the container to a second position adjacent to the outer surface of the container.

8. The purification method of claim 7, wherein, The actuator includes a pneumatic or hydraulic cylinder configured to move the plurality of magnetic field generating elements between the first position and the second position.

9. The purification method of claim 7 or 8, wherein, The container has an open end and a closed end, and a longitudinal axis extending between the open end and the closed end, wherein the actuator is configured to move the plurality of magnetic field generating elements in a direction not parallel to the longitudinal axis when the plurality of magnetic field generating elements are moved from the first position to the second position.

10. The purification method of claim 1, wherein, Moving the plurality of magnetic field generating elements simultaneously toward the container to apply the magnetic field and keep the magnetic beads abutting the inner surface of the container includes arranging a first permanent magnet and a second permanent magnet on opposite sides of the container.

11. The purification method of claim 1, wherein, Moving the plurality of magnetic field generating elements simultaneously toward the container to apply the magnetic field and keep the magnetic beads abutting the inner surface of the container includes arranging a plurality of permanent magnets at equal intervals around the perimeter of the container.

12. The purification method of claim 11, wherein, Each of the plurality of permanent magnets has a maximum magnetic pull force equal to or greater than 100 N.

13. The purification method according to claim 1, wherein, The target substance is a biomolecule.

14. The purification method of claim 1, wherein, The container has a volume equal to or greater than 0.5 L.

15. A purification system, comprising: A first container, configured to initially contain a mixture and magnetic beads for separating the target substance from the remainder of the mixture; A second container, which has a smaller volume than the first container; A plurality of magnetic field generating elements are arranged to surround the outer surface of the second container; A first pump, configured to transfer the mixture and the magnetic beads from the first container to the second container; A second pump, configured to remove the mixture from the second container; and The plurality of magnetic field generating elements can move simultaneously toward the second container to magnetically attract the magnetic beads to keep the magnetic beads against the inner surface of the second container, while the second pump removes the mixture from the second container.

16. The purification system of claim 15, wherein the second pump is configured to remove the mixture from the second container, while the first pump delivers the mixture and the magnetic beads from the first container to the second container, thereby enabling continuous flow of the mixture through the second container.

17. The purification system of claim 15 or 16, wherein the plurality of magnetic field generating elements are movable relative to the second container between a first position away from the second container and a second position adjacent to the outer surface of the second container.

18. The purification system of claim 17, wherein the plurality of magnetic field generating elements includes a first magnetic field generating element and a second magnetic field generating element, the first magnetic field generating element being configured to move toward and away from the second container along a first fixed path, and the second magnetic field generating element being configured to move toward and away from the second container along a second fixed path.

19. The purification system of claim 18, wherein the first fixed path is a straight line and aligned with a first imaginary radial line emanating from the longitudinal axis of the second container, and the second fixed path is a straight line and aligned with a second imaginary radial line emanating from the longitudinal axis of the second container.

20. The purification system of claim 17, further comprising an actuator configured to move the plurality of magnetic field generating elements from the first position to the second position.

21. The purification system of claim 20, wherein the actuator comprises a pneumatic cylinder or a hydraulic cylinder.

22. A purification method, comprising: Add the mixture, including the target substance, to the first container; Add the magnetic beads to the first container; The target substance is separated from the remaining substances in the mixture in the first container by means of the magnetic beads, wherein the target substance is temporarily bound to the magnetic beads; A first volume of mixture and a first number of magnetic beads are transferred from the first container to a second container, wherein the volume of the second container is smaller than the volume of the first container; Multiple magnetic field generating elements are arranged around the second container, and the multiple magnetic field generating elements are simultaneously moved toward the container to apply a magnetic field to keep the first number of magnetic beads in contact with the inner surface of the second container; The first volume of mixture is removed from the second container while the magnetic field keeps the first number of magnetic beads against the inner surface of the second container; The second volume of mixture and the second number of magnetic beads are transferred from the first container to the second container; and The second volume of mixture is removed from the second container while the magnetic field keeps the first and second number of magnetic beads against the inner surface of the second container.

23. The purification method of claim 22, wherein, Removing the first volume of mixture from the second container includes removing the first volume of mixture from the second container while simultaneously transferring the second volume of mixture and the second number of magnetic beads from the first container to the second container.

24. The purification method of claim 23, comprising adding a rinsing solution to the second container after removing the second volume of the mixture from the second container, and subsequently removing the rinsing solution from the second container while the magnetic field holds the first and second number of magnetic beads against the inner surface of the second container.

25. The purification method of claim 24, comprising removing the magnetic field after adding the rinsing solution to the second container, such that the first and second number of magnetic beads are freely dispersed in the rinsing solution, and subsequently reapplying the magnetic field during removal of the rinsing solution from the second container to keep the first and second number of magnetic beads abutting against the inner surface of the second container.

26. The purification method according to any one of claims 22 to 25, comprising: After the second volume of mixture is removed from the second container, eluent is added to the second container to elute the target substance bound to the first and second number of magnetic beads, and then the eluent and the target substance are removed from the second container while the first and second number of magnetic beads are held against the inner surface of the second container by the magnetic field.

27. The purification method of claim 26, comprising removing the magnetic field after adding the eluent to the second container, such that the first and second number of magnetic beads are freely dispersed in the eluent, and subsequently reapplying the magnetic field during removal of the eluent and the target substance from the second container to keep the first and second number of magnetic beads abutting against the inner surface of the second container.

28. The purification method according to claim 22, wherein, Transferring the first volume of mixture and the first number of magnetic beads from the first container to the second container includes pumping the first volume of mixture and the first number of magnetic beads via an automatic pump.

29. The purification method according to claim 28, wherein, Removing the combined first and second volumes of the mixture from the second container includes pumping the combined first and second volumes of the mixture from the second container via the automatic pump.

30. The purification method according to claim 22, wherein, Moving the plurality of magnetic field generating elements simultaneously toward the container to apply the magnetic field to keep the first number of magnetic beads abutting the inner surface of the second container includes: activating an actuator to move the plurality of magnetic field generating elements from a first position away from the second container to a second position adjacent to the outer surface of the second container.

31. The purification method of claim 30, wherein, The actuator includes a pneumatic or hydraulic cylinder configured to move the plurality of magnetic field generating elements between the first position and the second position.

32. The purification method according to claim 30 or 31, wherein, The second container has an open end and a closed end, and a longitudinal axis extending between the open end and the closed end, wherein the actuator is configured to move the plurality of magnetic field generating elements in a direction not parallel to the longitudinal axis when the plurality of magnetic field generating elements are moved from the first position to the second position.

33. The purification method according to claim 22, wherein, Moving the plurality of magnetic field generating elements simultaneously toward the container to apply the magnetic field to keep the first and second number of magnetic beads abutting the inner surface of the second container includes: positioning the first permanent magnet and the second permanent magnet on opposite sides of the container.

34. The purification method of claim 22, wherein, Applying the magnetic field to keep the first and second number of magnetic beads abutting the inner surface of the second container includes arranging a plurality of permanent magnets at equal intervals around the perimeter of the second container.

35. The purification method according to claim 34, wherein, Each of the plurality of permanent magnets has a maximum magnetic pull force equal to or greater than 100 N.

36. The purification method according to claim 22, wherein, The target substance is a biomolecule.

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