Separation system for purifying a biomolecule solution

Abstract

The present disclosure relates to a separation arrangement (100) for purifying a biomolecule solution (2), the separation arrangement (100) comprising: a container (10) for accommodating the biomolecule solution (2); magnetic particles (4) configured to be mixed with the biomolecule solution (2) inside the container (10) to bind a target molecule; a permanent magnet device (20) arranged exterior of the container (10); and an actuator device (30) arranged to move the container (10) and / or permanent magnet device (20) in relation to each other between a first and a second position (P1, P2); wherein the container (10) and the permanent magnet device (20) are positioned closer to each other in the second position (P2), such that the magnetic field of the permanent magnet device (20) captures and holds the magnetic particles (4) inside the container (10), whereby separation of target molecules from the remainder of the biomolecule solution (2) is enabled.

Description

[0001] Separation system for purifying a biomolecule solution

[0002] TECHNICAL FIELD

[0003] The present disclosure relates to a separation arrangement for purifying a biomolecule solution and a method for purifying a biomolecule solution by means of such separation arrangement. More specifically, the present disclosure relates to a separation arrangement for purifying a biomolecule solution and a method for purifying a biomolecule solution as defined in the introductory parts of the appended claims.

[0004] BACKGROUND ART

[0005] The process of purifying biomolecule solutions may nowadays comprise using magnetic beads binding a target molecule. For example, nucleic acid purification with magnetic beads relies on using magnetic beads with a coating that can bind nucleic acids reversibly by simply adjusting buffer conditions. This type of magnetic separation in molecular biology uses superparamagnetic beads and a magnetic field to provide a simple and reliable method of purifying various types of biomolecule. As an example, the target molecule first binds to the appropriately-coated magnetic beads. Next, an external magnetic field is applied, which attracts the beads to the outer edge of the container and immobilizes them. The magnetic beads with the bound target molecules typically remain immobilized during washing steps while contaminants are removed. Subsequently, elution buffer is added, which releases the target molecules from the immobilized beads and a purified sample, ready for quantitation and analysis, is achieved.

[0006] Magnetic beads typically incorporate small particles of iron oxides, such as magnetite (Fe3O4), which give them the superparamagnetic properties. The surface of the bead is modified to minimize non-specific binding of proteins. The bead surface is key to the functionality of magnetic beads. There are a wide range of surface chemistries available to suit any given application: carboxyl- and silica-coated magnetic beads for generic, nonspecific binding based on buffer conditions; oligo(dT)-coated beads for binding mRNA; and streptavidin-coated beads for binding biotinylated samples. With an appropriate bead 1

[0007] Cormderma " Company Proprietarysurface coating and optimized conditions, the target molecule will selectively bind to the magnetic beads, leaving contaminants in the solution. The purified target molecule can then be used directly in molecular biology analyses and applications.

[0008] Separation by using magnetic beads reduces the need for purification steps prior to the separation and minimizes the stress on the target molecules. However, know techniques are cumbersome and are not configured for large scale purification.

[0009] SUMMARY

[0010] It is an object of the present invention to provide a separation arrangement for purifying a biomolecule solution, which solves or alleviates at least some of the challenges with traditional separation arrangements in the pharma and biotech industry.

[0011] It is an object of the present invention to provide a separation arrangement and a method for purifying a biomolecule solution, which is efficient and reliable and which allows large volumes of biomolecule solution to be purified.

[0012] According to a first aspect of the present disclosure, there is provided a separation arrangement for purifying a biomolecule solution. The separation arrangement comprises: a container for accommodating the biomolecule solution; magnetic particles configured to be mixed with the biomolecule solution inside the container to bind a target molecule; a permanent magnet device arranged exterior of the container; and an actuator device arranged to move the container and / or permanent magnet device in relation to each other between a first and a second position; wherein the container and the permanent magnet device are positioned closer to each other in the second position, such that the magnetic field of the permanent magnet device captures and holds the magnetic particles inside the container, whereby separation of target molecules from the remainder of the biomolecule solution is enabled. When the magnetic particles with the target molecules are captured and held still by means of the permanent magnet device, the impurified remainder of the biomolecule solution can be discharged and clean elution buffer may be added to achieve a purified biomolecule solution comprising the target molecules.The permanent magnet device comprises at least one permanent magnet capable of maintaining its magnetization without the need for an external magnetic field. Commonly known separation arrangements typically use electromagnets, which are activated and deactivated to control the movability of the magnetic particles inside the container. A permanent magnet as used in the present invention is smaller and more lightweight than an equally strong electromagnet. The separation arrangement according to the invention can this way be smaller and more lightweight and thus be more user friendly. Furthermore, electromagnets require cooling and by using a permanent magnet device the separation process will comprise less steps and the arrangement will comprise fewer components.

[0013] The separation arrangement is configured to allows lOOmL to 50L of biomolecule solution to be purified.

[0014] The container and / or the permanent magnet device are movably arranged in relation to each other, such that the relative position between the container and the permanent magnet device can be changed. This way, it is controlled whether the magnetic field of the permanent magnet device attracts the magnetic particles in the container or not. In the first position, the distance between the container and the permanent magnet device is such that the magnetic field of the permanent magnet does not attract the magnetic particles inside the container. In the first position, the magnetic forces on the magnetic particles are thus deactivated and the magnetic particles are movable inside the container. The first position thus corresponds to the magnetic field being deactivated. In the second position, the distance between the container and the permanent magnet device is such that the magnetic field of the permanent magnet attracts and immobilizes the magnetic particles. Thus, in the second position, the magnetic particles will be retained against the inner wall of the container adjacent the permanent magnet device. In the second position, the magnetic forces on the magnetic particles are activated and the magnetic particles are not able to move inside the container. The second position thus corresponds to the magnetic field of the permanent magnet device being activated.

[0015] 3

[0016] Conridentia " Company ProprietaryThe biomolecule solution may be referred to as an IVT feed, such as mRNAfeed. However, the herein-mentioned separation arrangement and method are also applicable on other biomolecule solutions for purification of for example AAV. The biomolecule solution in the container before starting the purification process may be referred to as a starting material or a sample, whereas the biomolecule solution achieved at the end of the purification process may be referred to as a purified biomolecule solution.

[0017] In one example, the actuator device comprises an inflatable pillow arranged between the container and the permanent magnet device, wherein the inflatable pillow is configured to be in an inflated state in the first position and in a deflated state in the second position. When the inflatable pillow is inflated, the container is elevated outside the reach of the magnetic field of the permanent magnet device. When the inflatable pillow is deflated, the container is lowered towards the permanent magnet device within reach of the magnetic field of the permanent magnet device. The container may be resting on the inflatable pillow. The inflatable pillow may be configured such that it is essentially flat and very thin when being deflated. This way, the deflated pillow has neglectable influence on the magnetic field and the capability of capturing and holding the magnetic particles. The inflatable pillow is typically connected to a high flow pneumatic circuit, enabling inflation and deflation in a few seconds. The pneumatic circuit may comprise an ejector and at least one valve unit for alternating between inflation and deflation of the pillow.

[0018] The actuator device may comprise a moveable support plate and a mechanical elevator mechanism arranged to move the support plate. In one example, the actuator device comprises a moveable support plate and a mechanical elevator mechanism arranged to move the support plate, wherein the support plate is arranged between the container and the permanent magnet device. The support plate may be moved by means of an elevator mechanism including controllable cylinders. The container may be resting on the rigid support plate and the elevator mechanism may be configured to displace the support plate between an elevated position corresponding to the first position, and a lowered position corresponding to the second position. Thus, in the first position, the elevator mechanism 4

[0019] Conridentia " Company Proprietaryis controlled to elevate the support plate and thus the container outside the reach of the magnetic field of the permanent magnet device. In the second position, the elevator mechanism is controlled to lower the support plate and the container towards the permanent magnet device within reach of the magnetic field of the permanent magnet device. The support plate is configured such that it has neglectable influence on the magnetic field and the capability of capturing and holding the magnetic particles. The mechanical elevator mechanism may comprise two connected rods, where each rod comprises cam elements at each end. The cam elements are connected to the support plate, such that rotary movement of the cam elements will move the support plate vertically. The actuator device may further comprise hydraulic, pneumatic or electrical cylinders connected to the mechanical elevator mechanism. It is to be understood that the mechanical elevator mechanism could be configured in multiple ways, as long as it is able to move the support plate vertically.

[0020] In one example, the permanent magnet device comprises a plate with a plurality of block magnets. The plate with the plurality of block magnets may be referred to as the magnetic plate. The magnetic plate may be arranged underneath the inflatable pillow or the support plate as discussed above. The magnetic plate may comprise a steel back plate and a plurality of N35 or N52 Neodymium block magnets arranged on the plate. As an example, the magnetic plate may comprise between 50-100 block magnets, preferably between 70-90 block magnets. In one example, the magnetic plate comprises 88 block magnets. Each block magnet may have a length of 40 millimeters, a width of 32 millimeters and a thickness of 10 millimeters. It is to be understood that the block magnets could have any dimensions and the magnetic plate could comprise any number of block magnets.

[0021] The plurality of block magnets may be arranged on the plate with alternating north and south poles facing in direction of the container. This way, a suitable magnetic field gradient is generated and the magnetic attraction forces between the magnets and the backplate are strong and holds the assembly together.In another example, the permanent magnet device comprises a rod magnet assembly with a plurality of diametrically magnetized rod magnets and the container comprises pockets configured to receive the rod magnets in the second position. The rod magnets are thus configured to be inserted into the pockets of the container in the second position. This way, the magnetic particles inside the container will be within reach for the magnetic field of the rod magnets and the magnetic particles will be captured and held against the container walls surrounding the rod magnets. In the first position, the distance between the rod magnet assembly and the container is such that the magnetic particles inside the container are out of reach for the magnetic field of the rod magnets. The rod magnet assembly may be arranged above the container. The rod magnets may thus be lowered into the pockets of the container when being moved to the second position. Alternatively, the container is elevated to the second position, such that the rod magnets are slid into the pockets of the container. The actuator device may be a moveable arm, a pneumatic cylinder or similar, connected to the rod magnet assembly and configured to move the rod magnet assembly towards and away from the container. Alternatively, the actuator device is some sort of elevating mechanism arranged underneath the container to move the container towards and away from the rod magnet assembly.

[0022] The diametrically magnetized rod magnets may be N52 magnets. The rod magnets have an elongated shape and the pockets in the container have a corresponding shape. The rod magnet assembly may comprise 5-30 rod magnets. Typically, the rod magnets and the corresponding pockets are evenly distributed over the container. The rod magnets and the corresponding pockets may be arranged in parallel rows with similar distances between all rod magnets and pockets.

[0023] The rod magnet assembly may be detachably connected to the actuator device and the rod magnet assembly may be configured to be detached from the actuator device in the second position, to allow movement of the container in relation to the actuator device. Thus, when the rod magnets are inserted into the pockets of the container in the second position, the rod magnet assembly is suitably detached from the actuator device and the 6

[0024] Cormderma " Company Proprietarycontainer and the rod magnet assembly may be gently rocked in order to facilitate capturing of the magnetic particles. When the actuator device is controlled to move the rod magnet assembly to the first position to let the magnetic particles loose inside the container, the rod magnet assembly is reattached to the actuator device before withdrawing the rod magnets from the pockets. Typically, it is ensured that the container and the rod magnet assembly are arranged in a horizontal position before lifting the rod magnets out from the pockets.

[0025] The container comprises an inlet arrangement for filling the container with fluid and a discharge arrangement for draining the container. The inlet arrangement comprises an inlet aperture and the discharge arrangement comprises an outlet aperture. The inlet arrangement is typically connected to an inlet pump for pumping fluid into the container. The discharge arrangement is typically connected to a discharge pump for pumping fluid out from the container.

[0026] The container may be a rigid container. The container may thus comprise rigid walls. The container may comprise a top wall, a bottom wall and side walls. With a rigid container, the shape of the container will be constant and there will be a predetermined amount of air at the top of the container when filling the container with liquid. This will facilitate filling and draining of the container during the purification process. The outlet aperture of the discharge arrangement may be arranged in a side wall of the container, adjacent the bottom wall. The discharge arrangement, and specifically the outlet aperture, arranged close to the bottom will facilitate draining of the container and the container will this way be essentially completely emptied. The inlet arrangement may be arranged in a side wall of the container, wherein the inlet arrangement is inclined upwards in direction towards the top wall of the container. In one example, the inlet arrangement is arranged adjacent a lower corner of the container and is directed towards an opposing upper corner of the container. With an inclined inlet arrangement, the fluid that is pumped into the container will be sprayed over essentially the entire inside of the container, such that any magneticparticles stuck on the top wall or in upper corners of the container will be rinsed away. The container may further comprise a passive vent valve in the top wall of the container.

[0027] Alternatively, the container comprises a flexible bag. The flexible bag may comprise a top side and a bottom side. The flexible bag typically rests on its bottom side during the purification process. A flexible bag container is easier and less costly to manufacture. Such container would also be more lightweight, which is an advantage. The container may be a so-called cell bag, which is commonly used for bioprocessing. The flexible bag may comprise a plastic film welded together, and the discharge arrangement may be arranged at the periphery of the flexible bag. The container may further comprise a guiding element arranged to guide the solution inside the flexible bag towards a collection area in fluid communication with the discharge arrangement. The guiding element may comprise an inclined weld at one of the edges of the flexible bag, leading the fluid inside the flexible bag towards a corner of the flexible bag. The discharge arrangement may comprise an outlet aperture in the plastic film and a discharge pipe arranged inside the flexible bag between the outlet aperture and the collection area. The discharge arrangement may be arranged at an end of the flexible bag, with the outlet aperture in the top side of the flexible bag. The fluid inside the flexible bag will be guided towards the collection area and subsequently be sucked out through the outlet aperture via the discharge pipe. This will provide an efficient draining of the container. The flexible bag container may further comprise a trap device inside the flexible bag, arranged to prevent magnetic particles from being discharged together with the fluid. Such trap device may comprise a magnet arranged to attract any magnetic particles that are about to be discharged through the discharge arrangement. The inlet arrangement with the inlet aperture may be arranged in the top side of the flexible bag.

[0028] In one example, the permanent magnet device comprises a magnetized cylinder and the container is configured to be positioned inside the magnetized cylinder in the second position, and outside the magnetized cylinder in the first position. Using a magnetized cylinder which surrounds the container in the second position will enable improved 8

[0029] Conridentia " Company Proprietaryseparation properties because it generates a relatively even gradient of the magnetic field intensity in the entire volume of the biomolecule solution in the container. Hence, the radial forces acting on the magnetic particles in the biomolecule solution are relatively even in the entire volume of the container when being arranged inside the magnetized cylinder. Also, the center of the container will this way be completely free of magnetic particles. The magnetized cylinder may be a four pole Halbach cylinder. The container may in this example comprise a flask or bottle with a cylindrical shape configured to fit inside the magnetized cylinder. The container may comprise a bottom and a top. The container may be arranged such that the bottom is first inserted into the magnetized cylinder. The arrangement may be configured such that the container has a horizontal position or a vertical position. The discharge arrangement of the container may be arranged at the bottom of the container. The actuator device may comprise a pneumatic or hydraulic cylinder configured to move the container into and out from the magnetized cylinder.

[0030] The separation arrangement may further comprise a mixing arrangement configured to move the container in the first position. The mixing arrangement will thus move the container and facilitate mixing inside the container. As an example, the mixing arrangement may be used for mixing the biomolecule solution and the magnetic particles and thereby facilitate the target molecules binding to the magnetic particles. Mixing may be performed multiple times during the purification process and will be further described with regard to the purification method as disclosed herein. The mixing arrangement may be configured to move the container by means of shaking, stirring, rolling or rocking the container. The mixing arrangement may further be configured to move the container in the second position.

[0031] In one example, the mixing arrangement comprises a rocker on which the container is configured to be arranged. The rocker may typically be used as mixing arrangement when the permanent magnet device comprises the magnetic plate or the rod magnet assembly as described above. The actuator device may be arranged on the rocker or it may be arranged above the rocker and the container. In the event that the actuator device 9

[0032] Conridentia " Company Proprietarycomprises an inflatable pillow or a mechanical elevating mechanism, the actuator device as well as the permanent magnet device may be arranged on the rocker both in the first position and in the second position. In the event that the permanent magnet device comprises the rod magnet assembly and the actuator device is arranged above the container, the permanent magnet device will be arranged on the rocker only in the second position. With a rocker, it is crucial that the weight of the container, permanent magnet device and the actuator device is not too high. The rocker may also be activated in order to gently rock the container when being in the second position and for example thereby facilitate capturing of the magnetic particles.

[0033] In the event that the permanent magnet device comprises a magnetized cylinder, the mixing arrangement may be configured to shake the container when being in the first position. Thus, the mixing arrangement may be configured to shake the container when it is outside the magnetized cylinder. The shaking movement may be in the longitudinal direction of the container, thus towards and away from the magnetized cylinder. The mixing arrangement may be configured to provide a pulsating motion of the container. The mixing arrangement may comprise an orbital shaker including a motor and an eccentric wheel connected to the motor.

[0034] According to some examples, the actuator device is connected to an electrical control unit configured to control the actuator device and thereby the movement of the container and / or permanent magnet device in relation to each other. By automating the purification process, efficiency is increased and a larger amount of biomolecule solution can be purified.

[0035] According to another aspect of the present invention, there is provided a method for purifying a biomolecule solution by means of the separation arrangement as disclosed herein. The method comprises the steps of: controlling the actuator device so that that the container and the permanent magnet device are in the first position in relation to each other; mixing the magnetic particles with the biomolecule solution inside the container tobind the target molecules; controlling the actuator device to move the container and / or permanent magnet device to the second position to capture and hold the magnetic particles; washing the biomolecule solution by discharging the remaining biomolecule solution and adding a wash buffer; and releasing the target molecules from the magnetic particles by adding an elution buffer and thereby achieve a purified biomolecule solution. The method may be performed by the electrical control unit of the separation arrangement. It is to be understood that features and advantages as described with regard to the separation arrangement aspect are also applicable on the method aspect of the present invention.

[0036] The biomolecule solution and the magnetic particles may be pumped into the container either together or separately. The magnetic particles may be added in the form of a slurry with magnetic particles. In one example, the slurry of magnetic particles is pumped into the container first, where after the actuator device is controlled to move the container and / or the permanent magnet device to the second position to capture and hold the magnetic particles. Air may then be pumped into the container to clear flow paths from magnetic particles and the remaining liquid of the slurry is discharged. The biomolecule solution is subsequently pumped into the container. Once both the biomolecule solution and the magnetic particles are inside the container, the actuator device is controlled to move the container and / or the permanent magnet device to the first position where the magnetic particles will be out of reach for the magnetic field of the permanent magnet device.

[0037] The step of mixing the magnetic particles with the biomolecule solution may involve rocking or shaking the container in the first position. Specifically, the step of mixing the magnetic particles and the biomolecule solution may be performed by controlling the mixing arrangement to move the container in the first position. This way, binding the target molecules on the magnetic particles will be facilitated.After the mixing, the container and / or the permanent magnet device are moved to the second position to capture and hold the magnetic particles containing the target molecules. Air may subsequently be pumped into the container to clear flow paths of any magnetic particles. Subsequently, washing is performed by discharging the remaining biomolecule solution and adding a wash buffer.

[0038] The washing step may comprise controlling the actuator device to move the container and / or the permanent magnet to the first position to release the magnetic particles after addition of the wash buffer; mixing the magnetic particles with the wash buffer; controlling the actuator device to move the container and / or the permanent magnet to the second position to capture and hold the magnetic particles; and discharging the wash buffer. Before discharging the wash buffer, the washing step may comprise pumping air into the container to clear flow paths of any magnetic particles. The washing step may be performed multiple time to ensure a proper washing of the target molecules on the magnetic particles. As an example, the washing step may be repeated three times. When the last wash buffer has been pumped out from the container, the elution buffer can be added. The mixing of the magnetic particles and the wash buffer may be performed by controlling the mixing arrangement to move the container.

[0039] The method may further comprise controlling the actuator device to move the container and / or the permanent magnet to the first position to release the magnetic particles; mixing the magnetic particles with the elution buffer; controlling the actuator device to move the container and / or the permanent magnet to the second position to capture and hold the magnetic particles; and discharging the purified solution including the target molecules. Thus, after the elution buffer has been pumped into the container, the container and / or the permanent magnet device may be moved to the first position and mixing may be performed to facilitate the release of the target molecules from the magnetic particles. The mixing of the magnetic particles and the elution buffer may be performed by controlling the mixing arrangement to move the container. The container and / or the permanent magnet device are subsequently moved to the second position tocapture and hold the magnetic particles where after the elution buffer containing the target molecules (the purified biomolecule solution) can be discharged. Before discharging the elution buffer, the method may comprise pumping air into the container to clear flow paths of any magnetic particles. The steps involved for releasing the target molecules and achieving a purified biomolecule solution may be repeated multiple times. It is to be understood that the discharged elution buffer containing the target molecules, should be saved when the process steps are repeated.

[0040] When the elution buffer / purified biomolecule solution has been pumped out from the container, the magnetic particles will still be captured inside the container. The magnetic particles may then be reused.

[0041] The method may further comprise a post separation washing step for washing the magnetic particles inside the container after the purified biomolecule solution has been discharged. This way, the magnetic particles will be cleaned and can be reused for any other type of separation or purification process. The post separation washing step may involve adding a sanitation liquid into the container; controlling the actuator device to move the container and / or the permanent magnet device to the first position; mixing the magnetic particles and the sanitation liquid; controlling the actuator device to move the container and / or the permanent magnet to the second position to capture and hold the magnetic particles; and discharging the sanitation liquid. The mixing of the magnetic particles and the sanitation liquid may be performed by controlling the mixing arrangement to move the container. Before discharging the sanitation liquid, the post separation washing step may comprise pumping air into the container to clear flow paths of any magnetic particles. The above-mentioned steps may be repeated multiple times. The post separation washing step may further comprise adding a wash liquid into the container; controlling the actuator device to move the container and / or the permanent magnet device to the first position; mixing the magnetic particles and the wash liquid; controlling the actuator device to move the container and / or the permanent magnet to the second position to capture and hold the magnetic particles; and discharging the washliquid. The mixing of the magnetic particles and the wash liquid may be performed by controlling the mixing arrangement to move the container. Before discharging the wash liquid, the post separation washing step may comprise pumping air into the container to clear flow paths of any magnetic particles.

[0042] According to another aspect of the present invention, there is provided a container for use in a separation arrangement as disclosed herein. The container is configured for accommodating a biomolecule solution and magnetic particles. The container may comprise an inlet arrangement for filling the container with fluid and a discharge arrangement for draining the container. The inlet arrangement may comprise an inlet aperture and the discharge arrangement may comprise an outlet aperture. The inlet arrangement is typically connected to an inlet pump for pumping fluid into the container. The discharge arrangement is typically connected to a discharge pump for pumping fluid out from the container.

[0043] The container may comprise a flexible bag. The flexible bag may comprise a top side and a bottom side. The container may comprise a plastic film welded together and a discharge arrangement arranged at the periphery of the container, wherein the container further comprises a guiding element arranged to guide the solution inside the container towards a collection area in fluid communication with the discharge arrangement. The container may be a so-called cell bag, which is commonly used for bioprocessing. The guiding element may comprise an inclined weld at one of the edges of the flexible bag, leading the fluid inside the flexible bag towards a corner of the flexible bag. The discharge arrangement may comprise an outlet aperture in the plastic film and a discharge pipe arranged inside the flexible bag between the outlet aperture and the collection area. The discharge arrangement may be arranged at an end of the flexible bag, with the outlet aperture in the top side of the flexible bag. The fluid inside the flexible bag will be guided towards the collection area and subsequently be sucked out through the outlet aperture via the discharge pipe. This will provide an efficient draining of the container. The flexible bag container may further comprise a trap device inside the flexible bag, arranged toprevent magnetic particles from being discharged together with the fluid in the event that all magnetic particles are not captured and held by the magnetic field of the permanent magnet device. Such trap device may comprise a magnet arranged to attract any magnetic particles that are about to be discharged through the discharge arrangement. The inlet arrangement with the inlet aperture may be arranged in the top side of the flexible bag.

[0044] The container may alternatively be a rigid container and may comprise rigid walls. The container may comprise a top wall, a bottom wall and side walls. With a rigid container, the shape of the container will be constant and there will be a predetermined amount of air at the top of the container when filling the container with liquid. This will facilitate filling and draining of the container during the purification process. The outlet aperture of the discharge arrangement may be arranged in a side wall of the container, adjacent the bottom wall. The discharge arrangement, and specifically the outlet aperture, arranged close to the bottom will facilitate draining of the container and the container will this way be essentially completely emptied. The inlet arrangement may be arranged in a side wall of the container, wherein the inlet arrangement is inclined upwards in direction towards the top wall of the container. In one example, the inlet arrangement is arranged adjacent a lower corner of the container and is directed towards an opposing upper corner of the container. With an inclined inlet arrangement, the fluid that is pumped into the container will be sprayed over essentially the entire inside of the container, such that any magnetic particles stuck on the top wall or in upper corners of the container will be rinsed away. The container may further comprise a passive vent valve in the top wall of the container.

[0045] The present disclosure will become apparent from the detailed description given below. The detailed description and specific examples disclose preferred embodiments of the disclosure by way of illustration only. Those skilled in the art understand from guidance in the detailed description that changes and modifications may be made within the scope of the appended claims.

[0046] BRIEF DESCRIPTIONS OF THE DRAWINGS

[0047] 15

[0048] Conridentia " Company ProprietaryThe above objects, as well as additional objects, features and advantages of the present disclosure will be more fully appreciated by reference to the following illustrative and nonlimiting detailed description of example embodiments of the present disclosure, when taken in conjunction with the accompanying drawings, of which:

[0049] Figures la-c schematically illustrate a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0050] Figure 2 schematically illustrates details of a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0051] Figures 3a-d schematically illustrate a detail of a separation arrangement for purifying a biomolecule solution according to examples of the present disclosure;

[0052] Figure 4a-b schematically illustrate a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0053] Figure 5 schematically illustrates details of a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0054] Figure 6a-b schematically illustrate a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0055] Figure 7 schematically illustrates details of a separation arrangement for purifying a biomolecule solution according to an example of the present disclosure;

[0056] Figures 8a-b schematically illustrate containers for use in a separation arrangement according to examples of the present disclosure; and

[0057] 16

[0058] Conridentia " Company ProprietaryFigure 9 illustrates a diagram of a method for purifying a biomolecule solution by means of the separation arrangement as disclosed herein.

[0059] DETAILED DESCRIPTION

[0060] The present disclosure will now be described with reference to the accompanying drawings, in which preferred example embodiments of the disclosure are shown. The disclosure may, however, be embodied in other forms and should not be construed as limited to the herein disclosed embodiments. The disclosed embodiments are provided merely to fully convey the scope of the disclosure to the skilled person.

[0061] It is to be understood that the terminology used herein is for purpose of describing particular embodiments only, and is not intended to be limiting. It should be noted that, as used in the specification and the appended claims, the articles "a", "an", "the", and "said" are intended to mean that there are one or more of the elements unless the context explicitly dictates otherwise. Thus, for example, reference to "a unit" or "the unit" may include several devices, and the like. Furthermore, the terms "comprising", "including", "containing" and similar wordings are intended to be open-ended transitional terms that do preclude the possibility of additional elements or steps.

[0062] Figures la-c schematically illustrate a separation arrangement 100 for purifying a biomolecule solution according to an example of the present disclosure. The separation arrangement 100 comprises a container 10 for accommodating the biomolecule solution 2 and magnetic particles 4 configured to be mixed with the biomolecule solution 2 inside the container 10 to bind a target molecule. The separation arrangement 100 also comprises a permanent magnet 20 device arranged exterior of the container 10.

[0063] Furthermore, the separation arrangement 100 comprises an actuator device 30 arranged to move the container 10 and / or permanent magnet device 20 in relation to each other between a first and a second position Pl, P2. The container 10 and the permanent magnet device 20 are positioned closer to each other in the second position P2 than in the firstposition Pl, such that the magnetic field of the permanent magnet device 20 captures and holds the magnetic particles 4 inside the container 10 in the second position P2. This way, in the second position P2, separation of target molecules from the remainder of the biomolecule solution 2 is enabled. When the magnetic particles 4 with the target molecules are captured and held still by means of the permanent magnet device 20, the impurified remainder of the biomolecule solution 2 can be discharged and clean elution buffer may be added to achieve a purified biomolecule solution comprising the target molecules.

[0064] Figure la shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the first position Pl in relation to each other and figure lb shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the second position P2 in relation to each other.

[0065] In this example, the actuator device 30 comprises an inflatable pillow 32 arranged between the container 10 and the permanent magnet device 20. The inflatable pillow 32 is configured to be in an inflated state in the first position Pl and in a deflated state in the second position P2. As seen in Figure la, when the inflatable pillow 32 is inflated, the container 10 is elevated outside the reach of the magnetic field of the permanent magnet device 20. As shown in Figure lb, when the inflatable pillow 32 is deflated, the container 10 is lowered towards the permanent magnet device 20 within reach of the magnetic field of the permanent magnet device 20. Thus, in the second position P2, the magnetic particles 4 will be captured at the bottom of the container 10, adjacent the permanent magnet device 20. While the magnetic particles 4 comprising the target molecules are held immobilized at the bottom of the container 10, the remaining solution can be discharged and washing of the magnetic particles and target molecules can be performed to eventually achieve a purified biomolecule solution comprising the target molecules.

[0066] The separation arrangement 100 may further comprise a housing 40 with an open top for accommodating the container 10, the actuator device 30 (inflatable pillow 32) and thepermanent magnet device 20. The container 10 may be resting on the inflatable pillow 32. The inflatable pillow 32 may be configured to, in the deflated state, have neglectable influence on the magnetic field and the capability of capturing and holding the magnetic particles 4. The inflatable pillow 32 is typically connected to a high flow pneumatic circuit (not shown), enabling inflation and deflation in a few seconds.

[0067] In this example, the permanent magnet device 20 comprises a plate 22 with a plurality of block magnets 24, which will be further described with regard to Figure 2.

[0068] As shown in Figure lc, the separation arrangement 100 may further comprise a mixing arrangement 50 configured to move the container 10 in the first position Pl. The mixing arrangement 50 will thus move the container 10 and facilitate mixing inside the container 10. As an example, the mixing arrangement 50 may be used for mixing the biomolecule solution 2 and the magnetic particles 4 and thereby facilitate the target molecules binding to the magnetic particles 4, or for mixing the magnetic particles 4 and a wash liquid etc. The mixing arrangement 50 may be configured to move the container 10 by means of shaking, stirring, rolling or rocking the container 10. The mixing arrangement 50 may further be configured to move the container 10 in the second position.

[0069] In this example, the mixing arrangement 50 comprises a rocker 52 on which the container 10 is arranged. The housing 40 comprising the container 10, permanent magnet device 20 and the inflatable pillow 32 is typically arranged on the rocker 52 as a complete unit. Rockers are commonly used when handling and controlling cell cultures, and the rocker 52 of the mixing arrangement 50 may be such a common rocker. Thus, no additional mixing arrangement 50 will be required during the purification process according to the present invention. The rocker 52 may be controlled to vary the speed (rocking cycles per minute), the tilting angle and the acceleration or motion. The rocker 52 is typically controlled to an essentially horizontal position before moving the container 10 and / or the permanent magnet device 20 between the first and second position Pl, P2.Figure lc also shows the separation arrangement 100 comprising an electrical control unit 200 configured to control the actuator device 30 and thereby the movement of the container 10 and / or permanent magnet device 20 in relation to each other.

[0070] Figure 2 schematically illustrates a permanent magnet device 20 of a separation arrangement 100 according to an example of the present invention. The permanent magnet device 20 may be used in a separation arrangement 100 as disclosed in Figures la, lb and lc. Thus, the permanent magnet device 20 comprises a plate 22 with a plurality of block magnets 24.

[0071] The plate 22 with block magnets 24 is configured to be arranged underneath the inflatable pillow 32. The plate 22 may comprise a steel back plate and the block magnets 24 may be N35 or N52 Neodymium block magnets 24 arranged on the plate 22. It is to be understood that the block magnets 24 could have any dimensions and the magnetic plate 22 could comprise any number of block magnets 24.

[0072] The plurality of block magnets 24 may be arranged on the plate 22 with alternating north and south poles facing in direction of the container 10 as shown in this figure. This way, a suitable magnetic field gradient is generated and the magnetic attraction forces between the block magnets 24 and the plate 22 are strong and holds the assembly together.

[0073] Figure 3a-d schematically illustrates details of a separation arrangement 100 according to examples of the present invention. The separation arrangement 100 may be configured as disclosed in Figure la-c except for the actuator device 30. In this example, the actuator device 30 comprises a moveable support plate 34 and a mechanical elevator mechanism 36 arranged to move the support plate 34 vertically, thus upwards and downwards. The moveable support plate 34 is arranged in the housing 40 and the housing 40 may thus comprise vertical slots allowing the moveable support plate 34 to move vertically inside the housing 40.Figures 3a-b show one example of the mechanical elevator mechanism 36 and figures 3c-d show another example of the mechanical elevator mechanism 36. The support plate 34 is arranged between the container 10 and the permanent magnet device (not shown). The support plate 34 is moved by means of the elevator mechanism 36 including controllable cylinders 39. The controllable cylinders 39 may be hydraulic, pneumatic or electrical cylinders connected to the mechanical elevator mechanism 36. The container 10 may be resting on the rigid support plate 34 and the elevator mechanism 36 may be configured to displace the support plate 34 between an elevated position corresponding to the first position Pl, shown in Figure 3a and Figure 3c, and a lowered position corresponding to the second position P2, shown in Figure 3b and Figure 3d. Thus, in the first position Pl, the elevator mechanism 36 is controlled to elevate the support plate 34 and thus the container 10 outside the reach of the magnetic field of the permanent magnet device 20. In the second position P2, the elevator mechanism 36 is controlled to lower the support plate 34 and the container 10 towards the permanent magnet device 20 within reach of the magnetic field of the permanent magnet device 20. The support plate 34 is configured such that it has neglectable influence on the magnetic field and the capability of capturing and holding the magnetic particles 4 in the second position P2. It is to be understood that in the event that the permanent magnet device 20 is arranged above the container 10, the elevator mechanism 36 is controlled to elevate or lift the support plate 34 and the container 10 towards the permanent magnet device 20 in the second position P2 and lower the support plate 34 and the container 10 away from the permanent magnet device 20 in position Pl.

[0074] The mechanical elevator mechanism 36 shown in Figures 3a-b comprises two connected rods 37, where each rod 37 comprises cam elements 38 at each end. The cam elements 38 are connected to the support plate 34, such that rotary movement of the cam elements 38 will move the support plate 34 linearly.

[0075] Figures 4a-b schematically illustrate a separation arrangement 100 for purifying a biomolecule solution according to an example of the present disclosure. The separationarrangement 100 comprises a container 10 for accommodating the biomolecule solution 2 and magnetic particles 4 configured to be mixed with the biomolecule solution 2 inside the container 10 to bind a target molecule. The separation arrangement 100 also comprises a permanent magnet 20 device arranged exterior of the container 10.

[0076] Furthermore, the separation arrangement 100 comprises an actuator device 30 arranged to move the container 10 and / or permanent magnet device 20 in relation to each other between a first and a second position Pl, P2. The container 10 and the permanent magnet device 20 are positioned closer to each other in the second position P2 than in the first position Pl, such that the magnetic field of the permanent magnet device 20 captures and holds the magnetic particles 4 inside the container 10 in the second position P2. This way, in the second position P2, separation of target molecules from the remainder of the biomolecule solution 2 is enabled. When the magnetic particles 4 with the target molecules are captured and held still by means of the permanent magnet device 20, the impurified remainder of the biomolecule solution 2 can be discharged and clean elution buffer may be added to achieve a purified biomolecule solution comprising the target molecules.

[0077] Figure 4a shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the first position Pl in relation to each other and figure 4b shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the second position P2 in relation to each other.

[0078] In this example, the permanent magnet device 20 comprises a rod magnet assembly 26 with a plurality of diametrically magnetized rod magnets 28 and the container 10 comprises pockets 11 configured to receive the rod magnets 28 in the second position P2. The diametrically magnetized rod magnets 28 may be N52 magnets. The rod magnets 28 have an elongated shape and the pockets 11 in the container have a corresponding elongated shape. The rod magnet assembly 26 may comprise 5-30 rod magnets 28. The rod magnets 28 are inserted into the pockets 11 of the container 10 in the second positionP2. This way, the magnetic particles 4 inside the container 10 will be within reach for the magnetic field of the rod magnets 28 and the magnetic particles 4 will be captured and held against the container walls surrounding the rod magnets 28.

[0079] The rod magnet assembly 26 may be configured to be arranged above the container 10 and be moved essentially vertically between the first and the second position Pl, P2. The actuator device 30 may comprise a moveable arm 38 connected to the rod magnet assembly 26 and configured to move the rod magnet assembly 26 towards and away from the container 10. The rod magnets 28 are thus in this example lowered into the pockets 11 of the container 10 when being moved to the second position P2.

[0080] The rod magnet assembly 26 may be detachably connected to the actuator device 30. Thus, in this example, the rod magnet assembly 26 may be detachably connected to the moveable arm 38, such that it can be detached from the moveable arm 38 in the second position P2. This way, movement of the container 10 in relation to the actuator device 30 is enabled. Thus, when the rod magnets 28 have been inserted into the pockets 11 of the container 10 in the second position P2, the rod magnet assembly 26 is detached from the moveable arm 38 and the container 10 and the rod magnet assembly 26 can be gently rocked, for example in order to facilitate capturing of the magnetic particles 4. Thus, in the second position P2, the rod magnet assembly 26 will rest on top of the container 10. When the actuator device 30 should be controlled to move the rod magnet assembly 26 to the first position Pl to let the magnetic particles 4 loose inside the container 10, the rod magnet assembly 26 is reattached to the moveable arm 38 before retracting the arm 38 and withdrawing the rod magnets 28 from the pockets 11.

[0081] The separation arrangement 100 may further comprise a mixing arrangement 50 configured to move the container 10 in the first position Pl as also described with regard to Figures la and lb. Similar to the separation arrangement 100 disclosed in Figures la and lb, the mixing arrangement 50 in this example comprises a rocker 52 on which the container 10 is arranged. The rocker 52 may be controlled to vary the speed (rocking cyclesper minute), the tilting angle and the acceleration or motion to achieve a desired mixing. When the rod magnet assembly 26 is detached from the moveable arm 38 of the actuator device 30, the rocker 52 can gently rock the container 10 and the rod magnet assembly 26 to facilitate capturing the magnetic particles 4 around the inserted rod magnets 28.

[0082] Figure 5 shows the separation arrangement 100 according to Figures 4a-b. The figure shows a cross-sectional view of the pockets 11 of the container 10, when the rod magnets 28 are inserted into the pockets 11 and thus when the container 10 and the permanent magnet device 20 are in the second position P2 in relation to each other.

[0083] Figures 6a-b schematically illustrate a separation arrangement 100 for purifying a biomolecule solution according to an example of the present disclosure. The separation arrangement 100 comprises a container 10 for accommodating the biomolecule solution 2 and magnetic particles 4 configured to be mixed with the biomolecule solution 2 inside the container 10 to bind a target molecule. The separation arrangement 100 also comprises a permanent magnet 20 device arranged exterior of the container 10.

[0084] Furthermore, the separation arrangement 100 comprises an actuator device 30 arranged to move the container 10 and / or permanent magnet device 20 in relation to each other between a first and a second position Pl, P2. The container 10 and the permanent magnet device 20 are positioned closer to each other in the second position P2 than in the first position Pl, such that the magnetic field of the permanent magnet device 20 captures and holds the magnetic particles 4 inside the container 10 in the second position P2. This way, in the second position P2, separation of target molecules from the remainder of the biomolecule solution 2 is enabled. When the magnetic particles 4 with the target molecules are captured and held still by means of the permanent magnet device 20, the impurified remainder of the biomolecule solution 2 can be discharged and clean elution buffer may be added to achieve a purified biomolecule solution comprising the target molecules.Figure 6a shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the first position Pl in relation to each other and figure 6b shows the separation arrangement 100 where the container 10 and the permanent magnet device 20 are in the second position P2 in relation to each other.

[0085] In this example, the permanent magnet device 20 comprises a magnetized cylinder 29 and the container 10 is configured to be positioned inside the magnetized cylinder 29 in the second position P2, and outside the magnetized cylinder 29 in the first position Pl. The magnetized cylinder 29 circumferentially surrounds the container 10 in the second position P2 and will enable improved separation properties because it generates a relatively even gradient of the magnetic field intensity in the entire volume of the biomolecule solution 2 in the container 10. Hence, the radial forces acting on the magnetic particles 4 in the biomolecule solution 2 are relatively even in the entire volume of the container 10 when being arranged inside the magnetized cylinder 29. Also, the center of the container 10 will this way be completely free of magnetic particles 4. The magnetized cylinder 29 may be a four pole Halbach cylinder.

[0086] The container 10 in this example comprises a flask or bottle with a cylindrical shape configured to fit inside the magnetized cylinder 29. The container 10 may comprise a top side 16 and a bottom side 18. The container 10 is typically arranged such that the top side 16 is connected to the actuator device 30 and such that the bottom side 18 is first inserted into the magnetized cylinder 29. The separation arrangement 100 is herein illustrated such that the container 10 has a vertical position but it is to be understood that the separation arrangement 100 may be configured such that the container has a horizontal position. The container 10 further comprises an inlet arrangement (not shown) for filling the container 10 and a discharge arrangement (not shown) for draining the container 10. The inlet arrangement is typically connected to an inlet pump (not shown) for pumping fluid into the container 10. The discharge arrangement is typically connected to a discharge pump (not shown) for pumping fluid out from the container 10. The discharge arrangement of the container 10 may be arranged at the bottom side 18 of the container 10.The actuator device 30 may comprise at least one pneumatic or hydraulic cylinder 39 configured to move the container 10 into and out from the magnetized cylinder 29.

[0087] The separation arrangement 100 further comprises a mixing arrangement 50 configured to shake the container 10 when being in the first position Pl. Thus, the mixing arrangement 50 may be configured to shake the container 10 when it is outside the magnetized cylinder 29. The mixing arrangement 50 is shown in more detail in Figure 7.

[0088] In Figure 7, the separation arrangement 100 as disclosed in Figures 6a-b is arranged horizontally. The shaking movement may be in the longitudinal direction of the container 10, thus towards and away from the magnetized cylinder 29. The mixing arrangement 50 may in this example comprise an orbital shaker 54 including a motor and an eccentric wheel connected to the motor.

[0089] Figures 8a-b schematically illustrate containers 10 for a separation arrangement 100 according to examples of the present disclosure. The containers 10 are suitable for a separation arrangement 100 as disclosed in Figure la, lb, 2 and 3a-d.

[0090] Figure 8a shows a container 10 comprising a flexible bag 60. The flexible bag 60 comprises an inlet arrangement 12 for filling the bag 60 with fluid and a discharge arrangement 14 for draining the bag 60. The inlet arrangement 12 comprises an inlet aperture 13 and the discharge arrangement 14 comprises an outlet aperture 15. The inlet arrangement 12 is typically connected to an inlet pump (not shown) for pumping fluid into the bag 60. The discharge arrangement 14 is typically connected to a discharge pump (not shown) for pumping fluid out from the bag 60.

[0091] The flexible bag 60 comprises a top side 16 and a bottom side 18. The flexible bag 60 typically rests on its bottom side 18 during the purification process. The flexible bag 60 may be a so-called cell bag, which is commonly used for bioprocessing. The flexible bag 60may comprise a plastic film 61 welded together, and the discharge arrangement 14 may be arranged at the periphery of the flexible bag 60. The bag 60 further comprises a guiding element 62 arranged to guide the solution 2 inside the flexible bag 60 towards a collection area 64 in fluid communication with the discharge arrangement 14. The guiding element 62 may comprise an inclined weld at one of the edges of the flexible bag 60, leading the fluid 2 inside the flexible bag 60 towards a corner of the flexible bag 60. The discharge arrangement 14 further comprises a discharge pipe 66 arranged inside the flexible bag 60 between the outlet aperture 15 and the collection area 64. The fluid inside the flexible bag 60 will be guided towards the collection area 64 and subsequently be sucked out through the outlet aperture 15 via the discharge pipe 66. This will provide an efficient draining of the flexible bag 60.

[0092] The outlet aperture 15 of the discharge arrangement 14 may be formed in the plastic film 61. The outlet aperture 15 may be arranged at the top side 16 of the flexible bag 60. The inlet arrangement 12 with the inlet aperture 13 may be arranged at the top side 16 of the flexible bag 60.

[0093] The flexible bag 60 may further comprise a trap device (not shown) inside the flexible bag 60, arranged to prevent magnetic particles 4 from being discharged together with the fluid. Such trap device may comprise a magnet arranged to attract any magnetic particles 4 that are about to be discharged through the discharge arrangement 14.

[0094] Figure 8b shows a container 10 being a rigid container 70. The container 70 comprises a top wall 72, a bottom wall 74 and side walls 76, wherein all walls are rigid. The rigid container 70 comprises an inlet arrangement 12 for filling the container 70 with fluid and a discharge arrangement 14 for draining the container 70. The inlet arrangement 12 comprises an inlet aperture 13 in a container wall and the discharge arrangement 14 comprises an outlet aperture 15 in a container wall. The inlet arrangement 12 is typically connected to an inlet pump (not shown) for pumping fluid into the container 70. Thedischarge arrangement 14 is typically connected to a discharge pump (not shown) for pumping fluid out from the container 70.

[0095] The outlet aperture 15 of the discharge arrangement 14 is here arranged in a side wall 76 of the container 70, adjacent the bottom wall 74. The discharge arrangement 14, and specifically the outlet aperture 15, arranged close to the bottom will facilitate draining of the container 70.

[0096] The inlet arrangement 12, and thus the inlet aperture 13, is also arranged in a side wall 76 of the container 70. The inlet arrangement 12 is inclined upwards in direction towards the top wall 72 of the container 70. The inlet arrangement 12 may be arranged adjacent a lower corner of the container 70 and is directed towards an opposing upper corner of the container 70. With an inclined inlet arrangement 12, the fluid that is pumped into the container 70 will be sprayed over essentially the entire inside of the container 70, such that any magnetic particles 4 stuck on the top wall 72 or in upper corners of the container 70 will be rinsed away. The container 70 may further comprise a passive vent valve 78 in the top wall 72 of the container 70.

[0097] Figure 9 shows a diagram of a method for purifying a biomolecule solution by means of a separation arrangement 100 as disclosed herein. The separation arrangement 100 may be configured as disclosed in any of the previous figures.

[0098] The method comprises the steps of: controlling slOl the actuator device 30 so that that the container 10 and the permanent magnet device 20 are in the first position Pl in relation to each other; mixing sl02 the magnetic particles 4 with the biomolecule solution 2 inside the container 10 to bind the target molecules; controlling sl03 the actuator device 30 to move the container 10 and / or permanent magnet device 20 to the second position P2 to capture and hold the magnetic particles 4; washing sl04 the biomolecule solution 2 by discharging the remaining biomolecule solution and adding a wash buffer; and releasing sl05 the target molecules from the magnetic particles 4 by adding an elution buffer andthereby achieve a purified biomolecule solution. The method may be performed by the electrical control unit of the separation arrangement 100.

[0099] The step of mixing sl02 the magnetic particles with the biomolecule solution 2 may involve rocking or shaking the container 10 in the first position. Specifically, the step of mixing sl02 the magnetic particles 4 and the biomolecule solution 2 may be performed by controlling the mixing arrangement 50 to move the container 10 in the first position Pl. This way, binding the target molecules on the magnetic particles 4 will be facilitated.

[0100] After the mixing sl02, the container 10 and / or the permanent magnet device 20 are moved to the second position P2 to capture and hold the magnetic particles 4 containing the target molecules. Air may subsequently be pumped into the container 10 to clear flow paths of any magnetic particles 4. Subsequently, the step of washing sl04 is performed by discharging the remaining biomolecule solution and adding a wash buffer.

[0101] The washing step sl04 may comprise controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the first position Pl to release the magnetic particles 4 after addition of the wash buffer; mixing the magnetic particles 4 with the wash buffer; controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the second position P2 to capture and hold the magnetic particles 4 again; and discharging the wash buffer. Before discharging the wash buffer, the washing step sl04 may comprise pumping air into the container 10 to clear flow paths of any magnetic particles 4. The washing step sl04 may be performed multiple time to ensure a proper washing of the target molecules on the magnetic particles 4. When the last wash buffer has been pumped out from the container 10, the releasing step 105 can be initiated by adding the elution buffer. The mixing of the magnetic particles 4 and the wash buffer may be performed by controlling the mixing arrangement 50 to move the container 10.

[0102] 29

[0103] Conridentia " Company ProprietaryThe step of releasing sl05 the target molecules from the magnetic particles 4 may further comprise controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the first position Pl to release the magnetic particles 4; mixing the magnetic particles 4 with the elution buffer; controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the second position P2 to capture and hold the magnetic particles 4 again; and discharging the purified solution 2 including the target molecules. Thus, after the elution buffer has been pumped into the container 10, the container 10 and / or the permanent magnet device 20 may be moved to the first position Pl and mixing may be performed to facilitate the release of the target molecules from the magnetic particles 4. The mixing of the magnetic particles 4 and the elution buffer may be performed by controlling the mixing arrangement 50 to move the container 10. The container 10 and / or the permanent magnet device 20 are subsequently moved to the second position P2 to capture and hold the magnetic particles 4 again, where afterthe elution buffer containing the target molecules (the purified biomolecule solution) can be discharged. Before discharging the elution buffer, the method may comprise pumping air into the container 10 to clear flow paths of any magnetic particles 4. The steps involved for releasing the target molecules and achieving a purified biomolecule solution may be repeated multiple times.

[0104] When the elution buffer / purified biomolecule solution 2 has been pumped out from the container 10, the magnetic particles 4 will still be captured inside the container 10. The magnetic particles 4 may then be reused.

[0105] The method may further comprise a post separation washing step sl06 for washing the magnetic particles 4 inside the container 10 after the purified biomolecule solution 2 has been discharged. This way, the magnetic particles 4 will be cleaned and can be reused for any other type of separation or purification process. The post separation washing step sl06 may involve adding a sanitation liquid into the container 10; controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the first position Pl; mixing the magnetic particles 4 and the sanitation liquid; controlling theactuator device 30 to move the container 10 and / or the permanent magnet device 20 to the second position P2 to capture and hold the magnetic particles 4 again; and discharging the sanitation liquid. The mixing of the magnetic particles 4 and the sanitation liquid may be performed by controlling the mixing arrangement 50 to move the container 10. Before discharging the sanitation liquid, the post separation washing step sl06 may comprise pumping air into the container 10 to clear flow paths of any magnetic particles 4. The post separation washing steps may be repeated multiple times.

[0106] The post separation washing step sl06 may further comprise adding a wash liquid into the container 10; controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the first position Pl; mixing the magnetic particles 4 and the wash liquid; controlling the actuator device 30 to move the container 10 and / or the permanent magnet device 20 to the second position P2 to capture and hold the magnetic particles 4 again; and discharging the wash liquid. The mixing of the magnetic particles 4 and the wash liquid may be performed by controlling the mixing arrangement 50 to move the container 10. Before discharging the wash liquid, the post separation washing step sl06 may comprise pumping air into the container 10 to clear flow paths of any magnetic particles 4.

[0107] The person skilled in the art realizes that the present disclosure is not limited to the embodiments described above. The person skilled in the art further realizes that modifications and variations are possible within the scope of the appended claims.

[0108] 31

[0109] Conridentia " Company Proprietary

Claims

CLAIMS:

1. A separation arrangement (100) for purifying a biomolecule solution (2), the separation arrangement (100) comprising:a container (10) for accommodating the biomolecule solution (2); magnetic particles (4) configured to be mixed with the biomolecule solution (2) inside the container (10) to bind a target molecule;a permanent magnet device (20) arranged exterior of the container (10); andan actuator device (30) arranged to move the container (10) and / or permanent magnet device (20) in relation to each other between a first and a second position (Pl, P2);wherein the container (10) and the permanent magnet device (20) are positioned closer to each other in the second position (P2), such that the magnetic field of the permanent magnet device (20) captures and holds the magnetic particles (4) inside the container (10), whereby separation of target molecules from the remainder of the biomolecule solution (2) is enabled.

2. The separation arrangement (100) according to claim 1, wherein the actuator device (30) comprises an inflatable pillow (32) arranged between the container (10) and the permanent magnet device (20), wherein the inflatable pillow (32) is configured to be in an inflated state in the first position (Pl) and in a deflated state in the second position (P2).

3. The separation arrangement (100) according to claim 1, wherein the actuator device (30) comprises a moveable support plate (34) and a mechanical elevator mechanism (36) arranged to move the support plate (34), wherein the support plate (34) is arranged between the container (10) and the permanent magnet device (20).

324. The separation arrangement (100) according to any one of the preceding claims, wherein the permanent magnet device (20) comprises a plate (22) with a plurality of block magnets (24).

5. The separation arrangement (100) according to claim 4, wherein the plurality of block magnets (24) are arranged on the plate (22) with alternating north and south poles facing in direction of the container (10).

6. The separation arrangement (100) according to any one of claims 1-3, wherein the permanent magnet device (20) comprises a rod magnet assembly (26) with a plurality of diametrically magnetized rod magnets (28) and the container (10) comprises pockets (11) configured to receive the rod magnets (28) in the second position (P2).

7. The separation arrangement (100) according to claim 6, wherein the rod magnet assembly (26) is detachably connected to the actuator device (30) and wherein the rod magnet assembly (26) is configured to be detached from the actuator device (30) in the second position (P2), to allow movement of container (10) in relation to the actuator device (30).

8. The separation arrangement (100) according to any one of the preceding claims, wherein the container (10, 70) comprises rigid walls (72, 74, 76).

9. The separation arrangement (100) according to any one of claims 1-8, wherein the container (10) comprises a flexible bag (60).

10. The separation arrangement (100) according to claim 9, wherein the flexible bag (60) comprises a plastic film (61) welded together and a discharge arrangement (14) arranged at the periphery of the flexible bag (60), wherein the flexible bag (60) further comprises a guiding element (62) arranged to guide the solution (2) inside33the flexible bag (60) towards a collection area (64) in fluid communication with the discharge arrangement (14).

11. The separation arrangement (100) according to claim 10, wherein the discharge arrangement (14) comprises an outlet aperture (15) in the plastic film (61), and a discharge pipe (66) arranged inside the flexible bag (60) between the outlet aperture (15) and the collection area (64).

12. The separation arrangement (100) according to any preceding claim, wherein the permanent magnet device (20) comprises a magnetized cylinder (29) and the container (10) is configured to be positioned inside the magnetized cylinder (29) in the second position (P2), and outside the magnetized cylinder (29) in the first position (Pl).

13. The separation arrangement (100) according to claim 12, wherein the actuator device (30) comprises a pneumatic or hydraulic cylinder (39) configured to move the container (10) into and out from the magnetized cylinder (29).

14. The separation arrangement (100) according to any one of the preceding claims, further comprising a mixing arrangement (50) configured to move the container (10) in the first position (Pl).

15. The separation arrangement (100) according to claim 14, wherein the mixing arrangement (50) comprises a rocker (52) on which the container (10) is configured to be arranged.

16. The separation arrangement (100) according to any one of the preceding claims, wherein the actuator device (30) is connected to an electrical control unit (200) configured to control the actuator device (30) and thereby the movement of the container (10) and / or permanent magnet device (20) in relation to each other.

17. A method for purifying a biomolecule solution (2) by means of the separation arrangement (100) according to any one of the preceding claims, the method comprising the steps of:- controlling (slOl) the actuator device (30) so that that the container (10) and the permanent magnet device (20) are in the first position (Pl) in relation to each other;- mixing (sl02) the magnetic particles (4) with the biomolecule solution (2) inside the container (10) to bind the target molecules;- controlling (sl03) the actuator device (30) to move the container (10) and / or permanent magnet device (20) to the second position (P2) to capture and hold the magnetic particles (4);- washing (sl04) the biomolecule solution (2) by discharging the remaining biomolecule solution and adding a wash buffer; and- releasing (sl05) the target molecules from the magnetic particles (4) by adding an elution buffer and thereby achieve a purified biomolecule solution (2).

18. The method according to claim 17, wherein the step of mixing (sl02) the magnetic particles (4) with the biomolecule solution (2) involves rocking or shaking the container (10) in the first position (Pl).

19. The method according to any of claims 17-18, wherein the washing step (sl04) further comprises controlling the actuator device (30) to move the container (10) and / or the permanent magnet device (20) to the first position (Pl) to release the magnetic particles (4) after addition of the wash buffer; mixing the magnetic particles (4) with the wash buffer; controlling the actuator device (30) to move the container (10) and / or the permanent magnet device (20) to the second position (P2) to capture and hold the magnetic particles (4); and discharging the wash buffer.

20. The method according to any of claims 17-19, wherein the release step (sl05) further comprises controlling the actuator device (30) to move the container (10)and / or the permanent magnet device (20) to the first position (Pl) to release the magnetic particles (4); mixing the magnetic particles (4) with the elution buffer; controlling the actuator device (30) to move the container (10) and / or the permanent magnet device (20) to the second position (P2) to capture and hold the magnetic particles (4); and discharging the purified solution (2) including the target molecules.

21. The method according to any of claims 17-20, further comprising a post separation washing step (sl06) for washing the magnetic particles (4) inside the container (10) after the purified biomolecule solution (2) has been discharged.

22. A container (10, 60, 70) for use in a separation arrangement (100) according to any one of claims 1-16, the container (10) being configured for accommodating a biomolecule solution (2) and magnetic particles (4).

23. The container (10, 60, 70) according to claim 22, wherein the container (10) comprises a flexible bag (60).

24. The container (10) according to claim 23, wherein the flexible bag (60) comprises a plastic film (61) welded together and a discharge arrangement (14) arranged at the periphery of the flexible bag (60), wherein the flexible bag (60) further comprises a guiding element (62) arranged to guide the solution (2) inside the flexible bag (60) towards a collection area (64) in fluid communication with the discharge arrangement (14).

25. The container (10) according to claim 24, wherein the discharge arrangement (14) comprises an outlet aperture (15) in the plastic film (61), and a discharge pipe (66) arranged inside the flexible bag (60) between the outlet aperture (15) and the collection area (64).

26. The container (10) according to claim 22, wherein the container (10) comprises a top wall (72), a bottom wall (74) and side walls (76), wherein all walls are rigid.

27. The container (10) according to claim 26, further comprising an inlet arrangement (12) in a side wall (76), wherein the inlet arrangement (12) is inclined upwards in direction towards the top wall (72) of the container (70).

Images