Purification container

Abstract

A purification container for chromatographic purification of liquids. The purification container includes a seat and a chromatography medium assembly including: a chromatography membrane to which a target component contained in a liquid that is supplied to the container and passes through the chromatography membrane is bound; and a holder configured to press the chromatography film against the seat portion. Also disclosed are a chromatographic purification system, a method of separating a target component from a liquid sample comprising a target component and a non-target component prior to a purification and elution step in a chromatographic system, and a chromatographic purification method of separating a particular target component from a sample liquid.

Description

Technical Field

[0001] The present invention relates to a purification container for chromatographically purifying one or more target components suspended or contained in a liquid, such as molecules, cells, viruses, etc. The present invention also relates to a chromatographic purification system, a chromatographic purification method for separating a target component from a liquid sample containing the target component and non-target components, and a chromatographic purification method for binding and eluting the target component in a centrifugal chromatography system and / or a pumping chromatography system. Background Art

[0002] Chromatography is a purification technique for separating target components and non-target components such as chemical compounds or biological compounds contained in a liquid. Chromatography relies on chromatographic media or adsorbents that adsorb certain components. Chromatographic media can take different forms, such as particles, pellets, fibers, felts, monoliths or membranes. Liquid containing one or more target components is supplied to a container such as a columnar container containing a chromatographic medium. The amount of liquid supplied depends on the size and application of the container. On a small scale, a relatively small amount of liquid is usually supplied, and on a larger scale, more liquid is supplied. When provided in batches or discontinuously, this discrete amount of liquid can be referred to as a "sample". In the initial separation step, when the liquid flows through the chromatographic medium, the target component will be combined with the adsorbent. Once the liquid passes through the chromatographic medium and unbound impurities are washed away, any bound impurities can be removed by supplying a washing buffer through the chromatographic medium to further purify the specific target component bound to the chromatographic medium. Subsequently, at least one specific target component can be removed from the chromatographic medium by a suitable elution solution, which can be a buffer, a solvent or an aqueous co-solvent mixture that optionally contains a buffer.

[0003] With known chromatographic purification processes, the binding capacity of the target component is generally low, which, combined with the low sample processing throughput, results in extended processing times. Existing processes have been improved by providing new chromatographic membranes that have a larger binding surface area than other known membranes. For example, membranes made of nanofibers, such as cellulose nanofibers, increase the binding capacity by providing a much larger binding surface area than other known materials and are therefore more suitable for use, for example, in the capture of lentiviral vectors (LVVs), where high recovery and flow rates are essential. One such material developed by the applicant is called This is an enhanced cellulose nanofiber material that enables a highly receptive surface area for viral vectors, cells, and other biogenic compounds, allowing significantly faster flow rates than conventional chromatography materials, thereby increasing processing efficiency.

[0004] However, it has been found that with certain chromatography media component types, not all liquid is able to pass through the chromatography membrane mounted within the container. This is due to a poor seal between the membrane and the container wall. The passage of liquid around the chromatography membrane means that not all target components bind to the membrane, which results in a loss of recovered material relative to the volume of liquid supplied, reducing overall efficiency.

[0005] Traditionally, chromatography membranes can be ultrasonically or thermally bonded in place within a container to prevent liquids from bypassing or circumventing the membrane. However, non-thermoplastic membranes cannot be sealed or bonded to containers in this way, so the problem of liquids bypassing chromatography membranes remains an issue. This problem is particularly acute when chromatography membranes use new materials such as cellulose nanofibers. Summary of the invention

[0006] According to the present invention, there is provided a container for chromatographically purifying one or more target components contained in a liquid, the container comprising a seat and a chromatographic medium assembly, the chromatographic medium assembly comprising: a chromatographic membrane to which the target components are bound; and a retainer configured to compress a portion of the chromatographic membrane between the retainer and the seat.

[0007] The retainer may be formed from a resiliently deformable material and may be friction fit into the container.

[0008] The chromatographic media assembly may further include a compression spacer element in contact with a portion of the chromatographic membrane such that the retainer directly or indirectly engages the compression spacer element to press the chromatographic membrane against the seat. Another compression spacer element may be in contact with an opposing surface of a portion of the chromatographic membrane that faces the seat, although the periphery of the chromatographic membrane may also be in direct contact with the seat.

[0009] The chromatographic media assembly may include a single chromatographic membrane. Alternatively, the chromatographic media assembly may include a plurality of chromatographic membranes, in which case the assembly includes a compression spacer element spaced between each chromatographic membrane such that the chromatographic membranes are spaced from each other.

[0010] Regardless of whether the assembly includes one membrane or a plurality of membranes, the compression spacer element may be located above the uppermost membrane and / or below the lowermost membrane.

[0011] The chromatographic media assembly may include a first porous membrane support located between the single membrane or the uppermost membrane and a portion of the holder. The first porous membrane support may be received in the recess in the holder.

[0012] The chromatography assembly may further include a second porous membrane support located between the single membrane or the lowermost membrane and a portion of the container. The second porous membrane support may be accommodated in a recess formed in the container radially inward from the seat.

[0013] The membrane or each membrane can be formed by a fiber material derived with a group that can interact with the target component. In particular, the membrane or each membrane can be made of a material comprising nanofibers. In particular, the membrane or each membrane can be formed by an electrospun mixed nanofiber mat, which can be made of a derivatized cellulose and a non-cellulose-based polymer. Before or after being assembled in the container, a membrane derivatization with a group that can interact with the target component can be performed. Suitable groups for derivatization include, but are not limited to, ion exchange groups, polar groups, chelating groups, hydrophobic groups, hydrogen bonding groups, mixed mode groups or affinity ligands.

[0014] The container may include a cylindrical portion defining a first opening at one end and a conical portion tapering toward a second opening at an opposite end. The chromatographic media assembly may be located in the cylindrical portion.

[0015] The conical portion may include ridges defining a liquid flow distribution passage between the ridges in a direction between the cylindrical portion and the second opening. The chromatographic media assembly may be located above the ridges and may or may not be in contact with an upper surface of the ridges.

[0016] The second opening may include a fitting for attaching a liquid conduit to enable liquid to flow into or out of the container through the second opening.

[0017] The fitting may include a locking element for connecting the fluid conduit to the container. The lock may be a Luer lock. Alternatively, the lock may include a threaded connection or any other convenient way of connecting the fluid conduit to the container.

[0018] In some embodiments, the container may include a closure that is removably mounted to the cylindrical portion to close the first opening. The closure may provide an impermeable sealing cap or a venting cap.

[0019] The closure may include a conduit or tube defining a passageway enabling sample liquid to flow through the closure.

[0020] The tube may include a fitting including a locking element for connecting the fluid conduit to the closure. The locking may be a Luer lock. Alternatively, the locking may include a threaded connection or any other convenient way of connecting the fluid conduit to the closure. This feature facilitates use of the container in a bidirectional fluid flow mode. In other words, fluid may be pumped into the container through the tube or pumped out of the container through the tube.

[0021] The closure member may include a body portion received in the first opening of the cylindrical portion and a head portion engaged with an end portion of the cylindrical portion.

[0022] The closure member is push fit into the cylindrical portion.

[0023] The closure may include a resilient seal located between the body portion and the cylindrical portion to retain the body portion within the first opening.

[0024] According to one aspect of the present invention, a chromatography purification system is provided, comprising a container according to the present invention and a centrifuge tube, wherein the container is inserted into the centrifuge tube, a sample liquid containing a target component is placed in the container, and a centrifugal force is applied by using a centrifuge device to achieve fluid flow through a chromatographic membrane. It will be apparent to those skilled in the art that the sample liquid, the wash buffer and the elution solution can also be passed through the chromatographic membrane using this method.

[0025] According to another aspect of the present invention, a chromatographic purification system is provided, comprising: a container according to the present invention; a fluid collection container comprising a fluid conduit for attachment to one of the openings in the container; and a device for generating a continuous fluid flow from the sample fluid collection container through the container.

[0026] The chromatographic purification system may include a sample fluid collector including a fluid conduit for attachment to another of the openings in the container to receive fluid flowing out of the container.

[0027] The means for generating a continuous fluid flow may comprise a fluid pump.

[0028] The fluid conduit of the fluid receiver and the fluid conduit of the fluid collector may each include a mating piece for connecting to a complementary mating piece on the container.

[0029] According to another aspect of the present invention, there is provided a method for separating a target component from a liquid containing the target component and non-target components, the method comprising the following steps:

[0030] providing a fluid container containing a source of sample fluid;

[0031] The liquid is supplied from the collection container to a container, the container includes a chromatographic medium component, the chromatographic medium component includes a chromatographic membrane, the chromatographic membrane can bind to the target component so that the target component is adsorbed, and the liquid passes through the chromatographic membrane to remove unbound impurities.

[0032] The step of supplying the liquid from the receiving container into the container may include continuously supplying the liquid so that the liquid flows into and through the container.

[0033] The container may include a cylindrical portion defining a first opening at one end and a conical portion tapering toward a second opening at an opposite end. The method may include continuously supplying the liquid into the container through the first opening or the second opening.

[0034] The container may include a closure member that is removably mounted in the cylindrical portion to close the first opening. The closure member may include a conduit connector and a tube for allowing liquid to flow through the closure member. The method may include the step of continuously supplying the liquid so that the liquid flows through the tube in the closure member.

[0035] The step of supplying the liquid to cause the liquid to flow continuously through the container includes supplying the liquid into the container through the second opening so that the liquid leaves the container through the first opening in the tube in the closure.

[0036] According to another aspect of the present invention, there is provided a chromatographic purification method for separating a specific target component from a sample liquid containing a target component and a non-target component, the method comprising:

[0037] In the separation step, the target component is separated from the sample liquid by the following steps:

[0038] Providing a liquid source of the liquid;

[0039] supplying the liquid from the liquid source into a container, the container comprising a chromatographic medium assembly, the chromatographic medium assembly comprising a chromatographic membrane capable of binding to the target component, such that the liquid and impurities pass through the chromatographic membrane and the target component binds to the chromatographic membrane;

[0040] Also included in the washing step is the removal of any impurities bound to the chromatographic membrane by the following steps:

[0041] supplying a wash buffer into the container;

[0042] forcing the wash buffer through the chromatographic membrane, and

[0043] The step of separating the specific target components bound to the chromatographic membrane is also included by eluting the specific target components bound to the chromatographic membrane by the following steps:

[0044] supplying an elution solution into the container; and

[0045] The elution solution is forced through the chromatographic membrane.

[0046] The step of forcing the wash buffer through the chromatographic membrane may comprise placing the container in a centrifuge device and operating the centrifuge device.Alternatively, the container may be connected to a pump and the wash buffer may be pumped through the chromatographic membrane.

[0047] The step of forcing the eluent solution through the chromatographic membrane may comprise placing the container in a centrifugal device and operating the centrifugal device.Alternatively, the container may be connected to a pump and the eluent solution may be pumped through the chromatographic membrane.

[0048] The separation step may include supplying the liquid into the container such that the liquid flows through the chromatographic membrane in a first direction.

[0049] The separating step may include supplying the liquid into the container so that the liquid continuously flows into and out of the container along the first direction.

[0050] The container may have a first opening and a second opening, the chromatographic media assembly being located between the first opening and the second opening, and the separating step may include supplying the liquid so that there is a constant flow into and out of the container through the first opening and the second opening.

[0051] The separating step may include, before supplying the liquid, connecting a fluid inlet conduit to one of the first opening and the second opening, and connecting a fluid outlet conduit to the other of the first opening and the second opening, so that a constant flow flows into and out of the container through the fluid inlet conduit, the fluid outlet conduit, the first opening, and the second opening.

[0052] The washing step may include forcing the wash buffer to pass through the chromatographic membrane in a direction opposite to a first direction in which the liquid is supplied into the container in the separating step.

[0053] The eluting step may include supplying the eluting solution into the container and forcing the eluting solution to pass through the chromatographic membrane in a direction opposite to a first direction in which the liquid is supplied into the container in the separating step. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Embodiments of the present invention will now be described, by way of example only, with reference to the following drawings, in which:

[0055] Figure 1 shows a side view of a container for a centrifugal chromatography system;

[0056] Figure 2 Shown along Figure 1 A cross-sectional view of the container taken along line aa;

[0057] Figure 3 for Figure 2 an enlarged view of a portion of the container (circled and marked B);

[0058] Figure 4 shows another cross-section of a container according to an embodiment of the present invention;

[0059] Figure 5 Shows Figure 4 A partially cutaway perspective view of the middle container;

[0060] Figure 6 for Figure 4 an enlarged view of a portion of the container shown, detailing the chromatographic media components;

[0061] Figure 7 shows a cutaway perspective view of a portion of a chromatography membrane and compression spacer elements on either side of the membrane as a retainer compresses the periphery of the chromatography membrane into position between compression spacer elements;

[0062] Figure 8 shows a perspective view of a compressed spacer element;

[0063] Fig. 9 A perspective view of a chromatography membrane is shown;

[0064] Fig.10 A perspective view of a retaining element is shown with a portion removed for clarity;

[0065] Fig.11 for Figures 4 to 6 a cutaway perspective view of the container shown, but with the membrane, compressed spacer element and retaining element shown in spaced or exploded form (i.e., not compressed) for ease of understanding;

[0066] Fig. 12A is a cross-sectional view of a container using a single membrane according to another embodiment;

[0067] Fig. 12B for Fig. 12A A partially cut-away perspective view of a container;

[0068] Fig.13 for Fig. 12B an enlarged view of a portion of the container shown;

[0069] Fig.14A For along Figure 4 or Fig. 12A Cross-sectional view taken along line BB;

[0070] Fig. 14B is a partially cut-away perspective view of a container without a chromatographic medium assembly;

[0071] Fig.15 is a cross-sectional view of a container according to any embodiment of the present invention, the container further comprising a closure member received within an upper end thereof;

[0072] Fig.16 shows two containers within tubes located in a fixed angle rotor of a centrifuge; and

[0073] Fig.17 An arrangement is shown in which a pump may be used to draw fluid from a reservoir through a container. DETAILED DESCRIPTION

[0074] refer to Figure 1 , shows a container 1 for a chromatography purification system to receive a liquid sample containing target components and non-target components. Fig.16 An example of such a system is shown and described in more detail below.

[0075] The target component is a chemical or biological component in a sample that is separated from the rest of the sample so that the target component can be measured and / or otherwise processed. The target biological component can be, for example, a protein, a glycoprotein, a lipoprotein, a protein conjugate, an antibody, a plasma protein, a protein fragment, a peptide, an amino acid, a nucleic acid, an oligonucleotide, a nucleotide, a carbohydrate, a lipid, a virus, a viral vector, a lentiviral vector (LVV), an adeno-associated virus (AAV), a measles virus, an exosome, or a cell or cell particle. The target chemical component can be a metal ion, a radionuclide, a small molecule drug and related metabolites or other organic, inorganic or synthetic chemical entities.

[0076] In the orientation shown in each of the figures, the container 1 has an upper end and a lower end. The container 1 has an upper cylindrical portion 2 and a lower conical portion 4, wherein the upper cylindrical portion 2 has an open end 3 at the upper end of the container 1, and the lower conical portion 4 gradually tapers from the bottom of the cylindrical portion 2 to the lower end of the container 1. A connector 5 extends from the tip of the conical portion 4 and defines an opening 6 that passes through the connector 5 for allowing a liquid sample to flow into or out of the container 1. See Fig.14A As shown in Figure 4 and a cross-sectional view taken along line BB in the embodiment of FIG. 12 and Fig. 14BThe partially cutaway perspective view of the container 1 shown, the conical portion 4 includes an integrally formed internal ridge 7 rising from the inner surface of the conical portion 4 to the cylindrical portion 2. Each ridge 7 is spaced equidistantly from its adjacent ridge 7 to form a liquid distribution passage 8 between the ridges. In the embodiment shown, there are six ridges 7, and therefore six distribution passages 8 (one between each pair of ridges 7). It should be understood that in other embodiments not shown, there may be more or less than six ridges 7. Each distribution passage 8 extends along the inner wall surface of the conical portion 4 between the top of the connector 5 of the container 1 and the lower end of the cylindrical portion 2. The ridge 7 has a flat upper surface 7a. The ridge / seat structure can be a separate component that fits firmly in the conical portion 4 of the container 1. Alternatively, the ridge 7 and / or the seat 12 can be integrally molded into the container 1 during the manufacturing process.

[0077] As shown in the figure, the connecting member 5 may include a fitting 9 at its lower end away from the conical portion 4, that is, at the lower end of the container 1 (see Figure 1 and Figure 2 ). The fitting 9 allows a liquid conduit, such as a rigid tube or a flexible tube, to be connected to the liquid conduit. Fig.16 The flexible conduit mentioned can be engaged with the connector 5 to ensure a secure engagement. The fitting 9 can be configured to enable the fluid conduit to be absolutely or securely connected to the connector 5, i.e. locked to or matched with the connector 5, so that the fitting 9 cannot be removed simply by pulling or due to pressurized flow through the conduit and the connector 5. Instead, disconnection is required to separate one from the other. For example, the fitting 9 can be a Luer lock, a threaded coupling, or another type of locking arrangement that releasably connects the fluid conduit to the connector 5.

[0078] It should be understood that an absolute connection and disconnection is not necessary and a push fit or any other type of connection may be used. However, by establishing a secure connection between the sample liquid conduit and the connector 5, liquid can be supplied into the container 1 through the opening 6 without the liquid conduit accidentally falling off the connector 5. It also allows the liquid to be supplied or fed into the container 1 through the opening 6 under pressure without the liquid conduit accidentally falling off the connector 5, which may occur if the connection relies solely on friction between the two parts, such as if a more common Luer slip type connection is used. Since the liquid can be supplied at high pressure, higher flow rates can be achieved.

[0079] like Figure 3 , Figure 6 and Fig.11 As can be seen most clearly in the figure, a recess 10 is formed in the container 1, which is close to the top of the conical portion 4 of the container 1 and faces the inside of the cylindrical portion 2. The base of the recess 10 can be formed by the upper surface 7a of the ridge 7 (see Figure 6 , Fig.11 and Fig. 14B ) defines, as described above, a distribution passage 8 in the cone 4 is formed between the ridges 7. A shoulder 11 facing the longitudinal axis of the container 1 defines the edge or wall of the recess 10, the upper surface of the shoulder 11 forming a peripheral support or seat 12 facing the upper end of the container 1.

[0080] The chromatographic medium assembly 13 is contained in the container 1 and comprises a lower porous support member 14 located in the recess 10 and supported on the upper surface 7a of the ridge 7 (see Figure 3 , Figure 6 and Fig.10 ) on the upper surface, thereby being surrounded by the shoulder 11. The depth of the recess 10 is at least equal to the thickness of the lower porous support member 14.

[0081] In some embodiments, Fig. 12A , Fig. 12B and Fig.13 As shown, the chromatographic medium assembly 13 may include a single membrane 15, which may be optionally derivatized with an adsorbent coating that includes, for example, ion exchange or hydrophobic groups, chelating groups, multimodal ligands, or affinity ligands that adsorb certain desired target components. However, as will be described in reference Figures 2 to 6 and Fig.11 As stated, it is contemplated that the chromatographic media assembly 13 most preferably includes a plurality of membranes 15 positioned in a stacked relationship.

[0082] It should be understood that in addition to the single membrane or multiple membranes 15, the chromatographic media assembly 13 may also include resin beads (not shown) adjacent to or between the membranes to supplement the purification.

[0083] In a single membrane embodiment, such as Fig. 12A , Fig. 12B and Fig.13 As shown, the compression spacer 16 may be located on either or both sides of the single membrane 15, and the compression spacer 16 is preferably in the form of an annular ring having flat upper and lower surfaces. Figure 8 An example of a compression spacer element 16 can be seen in FIG. 1 , which has flat upper and lower surfaces 16a. Thus, the peripheral regions of the upper and lower surfaces of the membrane 15 are in contact with the flat surfaces 16a of the compression spacer element 16. However, in this single membrane construction, one or both of the compression spacer elements 16 may be omitted, in which case the retaining element (see below) would directly contact the upper surface of the membrane 15 around the periphery of the membrane 15, and the lower surface of the membrane 15 would also directly contact the seat 12 around the periphery of the seat.

[0084] like Figures 1 to 6 and Fig.11As shown, if the chromatographic media assembly 13 includes a plurality of membranes 15 positioned in a stacked relationship, each membrane 15 is spaced apart by a compressive spacer element 16 such that each membrane 15 is spaced apart from an adjacent membrane 15. In addition, the compressive spacer element 16 may be located above the uppermost membrane 15 and / or below the lowermost membrane 15. Figures 1 to 6 As shown, when the stack of membranes 15 together with their respective compression spacer elements 16 are held in place within the container 1, the stack of membranes 15 together with their respective compression spacer elements 16 are arranged and firmly held in place and abut against the seat 12 of the shoulder 11 at an upper position of the lower porous support member 14 (disposed in the recess 10). The membranes 15 extend radially outward beyond the diameter of the lower porous support member 14 so that the outer periphery of the membranes 15 is supported by the seat 12 extending radially outward from the upper end of the shoulder 11. In order to provide good support and improved sealing performance, it should also be noted that the width "W" (see FIG. 1 ) of the compression spacer element 16 in the radial direction is Figure 8 ) is greater than the radial width of the seat 12, so that the compression spacer element 16 extends radially inwardly beyond the recess 10 accommodating the lower porous support member 14. The number of membranes 15 is limited only by the size of the container 1.

[0085] Figure 7 1 shows a close-up cutaway perspective view of a portion of the peripheral region of a membrane 15, wherein the peripheral region of the membrane 15 is sandwiched between a pair of annular compression spacer elements 16. The thickness of each compression spacer element 16 is greater than the thickness of the membrane 15. In some embodiments, as Figure 7 As shown by the arrow marked "a" in FIG. 1 , the thickness of each compression spacer element 16 may be between 0.5 mm and 1 mm; Figure 7 The thickness of the film 15 shown at the arrow marked "b" in the figure may be between 0.01 mm and 0.4 mm.

[0086] When the retaining element 18 (see below) is in position within the container 1 so that the retaining element pushes the chromatographic medium assembly 13 against the seat 12, the peripheral region of the membrane 15 or each of the plurality of membranes 15 is compressed between the compression spacer elements 16. The compression of the membranes 15 closes the pores or gaps between the fibers of the chromatographic membrane 15, thereby making the peripheral region of each membrane 15 less permeable or at least semi-permeable to liquid than the more central, uncompressed region. The passage of liquid through the peripheral edge of the chromatographic membrane 15 is prevented or minimized, thereby preventing or minimizing liquid from bypassing the outside of the membrane 15 between the peripheral edge of the membrane 15 and the inner surface of the cylindrical portion 2 of the container 1. As shown in FIG. Figure 7 As shown, in the absence of any compressive force, the thickness of the combined compression spacer-membrane-compression spacer unit can be 20% to 90% thinner than the compression spacer-membrane-compression spacer.

[0087] An upper porous support 17 in the form of a flat disc is located above the uppermost membrane 15, or if only a single membrane is provided, above the membrane 15. If multiple membranes 15 are provided, each membrane may be formed of the same or different materials, and each membrane may have the same shape and size. The porous support members 14, 17 may be made of a material that is more rigid than the material of the membrane 15 to help maintain the flat shape of the membrane 15 and prevent any deformation caused by the weight or force of the liquid supplied to the container 1. The porous support member 14, 17 or each porous support member may contact the corresponding uppermost or lowermost membrane 15, but one or both of the porous support members 14, 17 may also be spaced from its adjacent membrane 15, or only lightly contact its adjacent membrane 15.

[0088] The membrane 15 is pushed or pressed against the seat 12 by a retaining member 18, which is shaped in a manner similar to that of a Fig.10 is best shown in the cutaway perspective view of Figure 2 , Figure 3 , Figures 4 to 6 , Fig. 12A , Fig. 12B and Fig.13 The position of the retaining member 18 in the container 1 is also shown. Fig.11 It can also be seen in the exploded diagram.

[0089] Preferably, the retainer 18 is in the form of an annular ring that is received in the cylindrical portion 2 of the container 1. The retainer 18 has a downward pressure engagement surface 19 that contacts the membrane 15, or if there are multiple membranes 15, the uppermost membrane 15. The pressure engagement surface 19 may be in direct contact with the membrane 15, or may apply pressure to the membrane 15 through a compression spacer element 16 received between the uppermost membrane 15 and the retainer 18. The pressure engagement surface 19 may include a notch or standoff (not shown) for enhancing the retention of the membrane 15, particularly when a single membrane 15 is used. Alternatively, the pressure engagement surface 19 may be smooth and contact the compression spacer element 16 or membrane 15 around its entire circumference. The retainer 18 also has an outer surface 20 that faces the inside of the cylindrical portion 2 and contacts the cylindrical inner surface of the cylindrical portion 2.

[0090] The retainer 18 is elastically deformable, and the size and dimensions of the retainer 18 are defined so that it is press-fit, friction-fit, or at least relatively tightly sliding fit within the cylindrical portion 2, wherein the surface 20 of the retainer engages with the wall of the cylindrical portion 2. The inner wall of the cylindrical portion 2 of the container 1 is generally smooth, but the surface may have a certain degree of roughness to increase the friction between the inner surface of the cylindrical portion 2 and the outer surface 20 of the retainer 18.

[0091] The upper surface 22 of the retainer 18 is chamfered in a direction extending downward and away from the inner surface of the cylindrical portion 2 of the container 1 toward the upper porous support member 14. The chamfered surface 22 helps to direct liquid away from the inner surface of the cylindrical portion 2 and toward a more central portion of the chromatographic membrane 15.

[0092] The retainer 18 may be formed of any elastically deformable material such as an elastomeric material, and is pushed downwardly along the cylindrical portion 2 from the opening 3 at the upper end of the cylindrical portion 2, with the surface 20 of the retainer 18 sliding against the inner surface of the cylindrical portion 2 until the retainer 18 contacts the uppermost membrane 15 or compression spacer element 16. Pushing the retainer 18 further into the cylindrical portion 2 causes the retainer 18 to apply pressure to the membrane 15 and any compression spacer element 16 to push the above-mentioned membrane and any compression spacer element against the seat 12. The friction between the retainer 18 and the inner surface of the cylindrical portion 2 holds the retainer 18 in place. In addition, the pressure of the retainer 18 against the membrane 15 and the compression spacer element 16 holds the chromatographic medium assembly 13 in place, pressing the assembly 13 downward against the seat 12 and preventing the assembly from shifting.

[0093] Retaining member 18 (such as Figure 2 , Figure 3 , Figure 6 , Fig.10 and Fig.11 The retainer 18 (most clearly shown in FIG. 1 ) may have a radially inwardly extending lip 23 spaced from the pressure engagement surface 19, the lip 23 forming a recess to receive and retain the upper porous support 14 between the radially inwardly extending lip 23 and the pressure engagement surface 19. Thus, the retainer 18 also locates the upper porous support 17 and holds it in place within the recess. The retainer 18 squeezes the membrane 15 and the compression spacer element 16 together between the pressure engagement surface 19 of the retainer 18 and the seat 12 of the container. The retainer 18 supports one or more membranes 15 within the container 1, thereby preventing movement of the membrane 15 that may affect performance and / or sealing.

[0094] Furthermore, as described above, the combined compression spacer element 16 and membrane 15 are squeezed and compressed between the retainer 18 and the seat 12 so that any spaces or holes between the fibers of the membrane 15 in the peripheral region between the compression spacer elements 16 are closed or semi-closed, thereby preventing or limiting the radial flow of liquid toward the outer periphery of the membrane 15. By preventing the liquid from moving radially outward to the outer periphery of the membrane 15, the liquid is prevented from bypassing the outside of the membrane 15.

[0095] Fig.15A portion of the upper end of a container 1 according to any of the previous embodiments is shown. In this embodiment, the container 1 is also provided with a closure 25, a portion of which is received in the open upper end of the cylindrical portion 2. The closure 25 includes a body portion 26, which contains a passage 27, and a sealing element 28, such as an elastic O-ring, is received in the passage 27. In other embodiments not shown, a polypropylene protrusion connected to or associated with the closure 25 can be provided to provide a seal between the closure 25 and the container 1.

[0096] The dimensions of the main body portion 26 of the closure 25 can be defined and adapted to be pushed into the cylindrical portion 2 of the container 1, so that the head 29 having a diameter greater than the diameter of the main body portion 26 is seated on the end surface of the cylindrical portion 2 of the container 1 and is used to prevent the closure 25 from moving further into the cylindrical portion 2 of the container 1.

[0097] The closure 25 includes a channel 30 defined by a channel wall 31 (such as a cylindrical or other shaped tube) that allows liquid to flow into or out of the container 1 through the closure 25. A liquid conduit can be connected to the closure 25 at the opening of the channel 30 to allow fluid to pass between the conduit and the channel 30 and through the closure 25. Once the liquid conduit is connected to the channel 30 of the closure 25, the container 1 is isolated from the atmosphere. In other embodiments, the closure can be a flat cover or a vented cover to allow air to pass and equalize the internal pressure of the container during centrifugation and the associated liquid volume reduction.

[0098] It should be understood that the closure 25 can be attached to the container 1 in a manner other than a push fit. For example, the closure 25 and the container 1 can have complementary threaded connections to allow the closure 25 to be screwed onto the container 1. The opening of the channel 30 of the closure 25 can also be provided with a fitting similar to the connector 5 provided at the end of the conical portion 4, so that the liquid conduit can be securely connected to the channel wall 31 on the closure 25, that is, a Luer lock connection can be adopted. This feature facilitates the use of the container 1 in a bidirectional fluid flow mode. In other words, the fluid can be pumped into the container 1 through the channel 30, or pumped out of the container 1 through the channel 30.

[0099] When in use, a liquid sample containing target components and non-target components is supplied to the container 1, so that the liquid sample flows through a portion of the chromatographic medium component 13. Optionally, the membrane 15 has an adsorbent coating, or is derived from an adsorbent coating, which contains, for example, ion exchange or hydrophobic groups, chelating groups, multimodal ligands or affinity ligands, and the adsorbent coating adsorbs certain desired target components, so that these target components are bound to the adsorbent.

[0100] According to one embodiment, Fig.17As shown, a chromatography media purification system 32 is provided. A first liquid conduit 33 may be a flexible tube having one end attached to a connector 5 and leading to a collection container 34 via a pump 35. A second liquid conduit 36 ​​may be a flexible tube having one end connected to a reservoir 37 containing a liquid supply and connected to a collection container 34 via a closure, such as a pump 35. Fig.15 The closure member 25 shown (but Fig.17 The second liquid conduit 33 is a liquid conduit 35 (not shown) connected to the other end of the upper end of the cylindrical portion 2 of the container 1. This arrangement allows the liquid to flow from the reservoir 37 into the container 1 along the second liquid conduit in a continuous manner in response to the operation of the pump 35, pass through the membrane 15 of the chromatographic medium assembly 13, and flow out of the container 1 through the first liquid conduit 33 and into the collection container 34. In this way, it is possible to separate the target component from a liquid amount having a volume much larger than the volume of the container 1. This is particularly advantageous in the case of a large amount of liquid containing a relatively small amount of the target component.

[0101] Although the pump 35 is positioned to draw fluid from the reservoir 37 through the container 1 , the pump 35 may also be positioned in the second fluid conduit 36 ​​to push fluid through the container 1 .

[0102] The liquid sample can be caused to flow through the container 1 in either direction. For example, the liquid can be supplied to the container 1 through the opening 6 in the connector 5 and leave the container 1 through the channel 30 in the closure 25, or enter the container 1 through the channel 30 of the closure 25 housed at the upper end of the cylindrical portion 2 and leave the container 1 through the opening 6 of the connector 5. By allowing the sample fluid to flow into the container 1 through the opening 6 in the connector 5, the sample fluid flows along the distribution passage 8 between the ridges 7 in the conical portion 4 and can be more evenly distributed before the sample fluid flows through the single membrane 15 or multiple membranes 15 of the chromatographic medium assembly 13.

[0103] System 32 may include a control unit (not shown) to enable a user to control the flow rate of liquid through container 1. Liquid collected in collection container 34 may be disposed of. Liquid conduit 33 may replace collection container 34 to simply discharge waste liquid, such as to a drain.

[0104] Once the initial separation or capture step is complete, a wash step may be performed in which a wash buffer is supplied to the container 1 to further purify a specific target component from all target components bound to the membrane 15. It is envisioned that the wash buffer will be supplied to the container 1 in the same direction as the direction of liquid flow in the separation step described above, although the wash buffer may also be supplied to the container 1 in the opposite direction. The wash step may involve removing any liquid conduits 33, 36 from the container 1 and placing the container 1 in a centrifuge 38 (see Fig.16 ), thereby applying centrifugal force to push or force the wash buffer through the membrane 15.

[0105] Fig.16 In simplified form, a bag-shaped device 39 ( Fig.16 The centrifugal device 38 (two of which are shown) is used to receive centrifugal tubes 40 having closed lower ends 41. The container 1 is slidably received at the upper open end of each centrifugal tube 40. When the container 1 is subjected to centrifugal force, due to the rotation of the centrifugal device 38 around the axis XX, the washing buffer placed in the container 1 will be forced to pass through the membrane 15 and be discharged through the connector 5, thereby being collected in the closed lower ends 41 of the centrifugal tubes 40.

[0106] It will be appreciated that the wash buffer may be forced through the membrane 15 by other means, such as using a pump.

[0107] The wash buffer is selected so that impurities are released into the wash buffer to leave one or more target components bound to the membrane 15. The wash step may be performed one or more times with the same or different wash buffers to release different target components, thereby separating and leaving specific target components still bound to the membrane 15. By ensuring that the wash buffer is allowed to flow through the container 1 in the opposite direction to the flow of the liquid sample through the container 1 during the separation phase, unwanted components, particularly unwanted particulate impurities, are more easily released from the membrane 15, leaving the target component of interest on the membrane 15.

[0108] Then, a final elution step can be performed by introducing an elution solution into the container 1, which is used to elute the desired target component from the membrane 15. As with the washing step, the container 1 is placed in a centrifuge and, with the help of the elution solution, the centrifuge is centrifuged in the same manner as described above and using the same method as described above. Fig.16 The associated centrifugal device 32 elutes specific target components from the membrane 15 by applying centrifugal force. The elution solution is preferably introduced into the container 1, thereby allowing the elution solution to flow through the container 1 in the same direction as the wash buffer, i.e., in the opposite direction to the direction in which the liquid sample flows through the container 1 during the separation stage.

[0109] It will be appreciated that the elution solution may be forced through the membrane by other means, such as using a pump, rather than using a centrifuge.

[0110] The above content is regarded as the explanation of the principle of the present invention only. In addition, since many modifications and changes will occur easily to those skilled in the art, it is not desirable to limit the present invention to the exact construction and operation shown and described, and therefore, all suitable modifications and equivalents may be adopted, which are within the scope of the present invention.

Claims

1. A container for chromatographically purifying one or more target components contained in a liquid, the container comprising a seat and a chromatographic medium component, the chromatographic medium component comprising: at least one chromatographic membrane, the target component being bound to the at least one chromatographic membrane; and a retainer configured to compress a portion of the at least one chromatographic membrane between the retainer and the seat.

2. The container according to claim 1, wherein The retainer is formed of a resiliently deformable material and is friction fit into the container.

3. The container according to claim 2, wherein: The chromatographic media assembly further includes a compression spacer element in contact with a portion of the chromatographic membrane such that the retainer directly or indirectly engages the compression spacer element to press the chromatographic membrane against the seat.

4. The container according to claim 3, comprising: Another compression spacer element is in contact with an opposing surface of a portion of the chromatography membrane facing the seat.

5. The container according to claim 1, wherein The chromatographic media assembly includes a single chromatographic membrane.

6. The container according to any one of claims 1 to 4, wherein The chromatographic media assembly includes a plurality of chromatographic membranes, and the assembly includes a compression spacer element spaced between each chromatographic membrane such that the chromatographic membranes are spaced apart from each other.

7. The container according to any one of claims 4 to 6, wherein The chromatographic media assembly includes a first porous membrane support positioned between the single membrane or the uppermost membrane and a portion of the retaining member.

8. The container according to claim 7, wherein: The holder includes a recess, and the first porous membrane support is accommodated in the recess.

9. A container according to claim 8, comprising a second porous membrane support located between the single membrane or the lowermost membrane and a portion of the container.

10. A container according to claim 9, comprising a recess formed in the container radially inwardly from the seat, wherein The second membrane support is accommodated in the recess.

11. A container according to any preceding claim, wherein: The or each membrane is formed from a fibrous material derivatised with groups capable of interacting with the target component.

12. A container according to claim 11 wherein the or each membrane is made from a material comprising nanofibres, such as an electrospun hybrid nanofibre mat made from derivatised cellulose and non-cellulose based polymers.

13. A container according to any preceding claim, comprising: a cylindrical portion defining a first opening at one end and a conical portion tapering toward a second opening at an opposite end, the chromatographic media assembly being located in the cylindrical portion.

14. The container according to claim 13, wherein The conical portion includes ridges defining a liquid flow distribution passage between the ridges in a direction between the cylindrical portion and the second opening.

15. The container according to claim 14, wherein The chromatographic media assembly is located above the ridge.

16. A container according to any one of claims 13 to 15, wherein The second opening includes a fitting for attaching a liquid conduit to enable liquid to flow into or out of the container through the second opening.

17. The container according to claim 16, wherein The fitting includes a locking element for connecting the fluid conduit to the container.

18. The container according to claim 17, wherein The lock comprises a Luer lock.

19. The container according to claim 17, wherein: The locking comprises a threaded connection.

20. A container according to any one of claims 13 to 19, comprising a closure removably mounted to the cylindrical portion to close the first opening.

21. The container according to claim 20, wherein The closure includes a tube defining a passageway that enables sample liquid to flow through the closure.

22. The container according to claim 21, wherein The tube includes a fitting including a locking element for connecting a fluid conduit to the closure.

23. The container according to claim 22, wherein The locking element may be a Luer lock or a threaded connection.

24. A container according to any one of claims 20 to 23, wherein The closure member includes a body portion received in the first opening in the cylindrical portion and a head portion engaged with an end of the cylindrical portion.

25. The container according to claim 24, wherein The closure member is push-fit into the cylindrical portion.

26. The container according to claim 25, wherein The closure member includes a resilient seal located between the body portion and the cylindrical portion to retain the body portion within the first opening.

27. A chromatographic purification system, comprising a container and a centrifugal device according to any one of claims 1 to 26, wherein the container is inserted into the centrifugal device so that a sample liquid containing a target component placed in the container flows through the centrifugal device and flows through the chromatographic medium under the action of centrifugal force.

28. A chromatography purification system comprising: A container according to any one of the preceding claims; a fluid container including a fluid conduit for attachment to one of said openings in said container; and means for producing a continuous flow of fluid from said fluid reservoir through said container.

29. The chromatographic purification system according to claim 28, comprising: A fluid collector includes a fluid conduit for attachment to the other of the openings in the container to receive fluid flowing from the container.

30. The chromatography system according to claim 28 or 29, wherein The means for generating a continuous fluid flow comprises a fluid pump.

31. The chromatography system according to claim 29 or 30, wherein The fluid conduit of the fluid receiver and the fluid conduit of the fluid collector each include a mating piece for connecting to a complementary mating piece on the container.

32. A method for separating a target component from a liquid containing the target component and non-target components, the method comprising the following steps: providing a fluid container containing a source of liquid; The liquid is supplied from the collection container to a container, the container includes a chromatographic medium component, the chromatographic medium component includes a chromatographic membrane, the chromatographic membrane can bind to the target component so that the target component is adsorbed, and the liquid passes through the chromatographic membrane to remove unbound impurities.

33. The method of claim 32, wherein: The step of supplying the liquid from the receptacle into the container includes continuously supplying the liquid so that the liquid flows into and through the container.

34. The method of claim 33, wherein: The container includes a cylindrical portion defining a first opening at one end and a conical portion tapering toward a second opening at an opposite end, and the method includes continuously supplying the liquid into the container through the first opening or the second opening.

35. The method of claim 33, wherein: The container comprises a closure removably mounted in the cylindrical portion to close the first opening, the closure comprising a conduit connector and a tube for allowing the liquid to flow through the closure, wherein the method comprises the step of continuously supplying the liquid to allow the liquid to flow through the tube in the closure.

36. The method of claim 34, wherein: The step of supplying the liquid to cause the liquid to flow continuously through the container includes supplying the liquid into the container through the second opening so that the liquid leaves the container through the first opening in the tube in the closure.

37. A chromatographic purification method for separating a specific target component from a sample liquid containing the target component and non-target components, the method comprising: In the separation step, the target component is separated from the sample liquid by the following steps: Providing a liquid source of the liquid; supplying the liquid from the liquid source into a container, the container comprising a chromatographic medium assembly, the chromatographic medium assembly comprising a chromatographic membrane capable of binding to the target component, such that the liquid and impurities pass through the chromatographic membrane and the target component binds to the chromatographic membrane; Also included in the washing step is the removal of any impurities bound to the chromatographic membrane by the following steps: supplying a wash buffer into the container; forcing the wash buffer through the chromatographic membrane, and The step of separating the specific target components bound to the chromatographic membrane is also included by eluting the specific target components bound to the chromatographic membrane by the following steps: supplying an elution solution into the container; and The elution solution is forced through the chromatographic membrane.

38. The method of claim 36, wherein: Forcing the wash buffer through the chromatographic membrane includes placing the container in a centrifuge device and operating the centrifuge device.

39. The method of claim 36, wherein: Forcing the wash buffer through the chromatographic membrane includes attaching the container to a pump and pumping the wash buffer through the container.

40. The method according to any one of claims 36 to 38, wherein: Forcing the elution solution through the chromatographic membrane includes placing the container in a centrifugal device and operating the centrifugal device.

41. The method according to any one of claims 36 to 38, wherein: Forcing the eluent solution through the chromatographic membrane includes attaching the container to a pump and pumping the eluent solution through the container.

42. The method according to any one of claims 36 to 40, wherein: The separation step includes supplying the liquid into the container and allowing the liquid to flow through the chromatographic membrane in a first direction.

43. The method of claim 41, wherein: The separating step includes supplying the liquid into the container so that the liquid continuously flows into and out of the container along the first direction.

44. The method of claim 40, wherein: The container has a first opening and a second opening, the chromatographic media assembly is located between the first opening and the second opening, and the separating step includes supplying the liquid so that there is a constant flow into and out of the container through the first opening and the second opening.

45. The method of claim 40, wherein: The separation step includes: before supplying the liquid, connecting a fluid inlet conduit to one of the first opening and the second opening, and connecting a fluid outlet conduit to the other of the first opening and the second opening, so that there is a constant flow into and out of the container through the fluid inlet conduit, the fluid outlet conduit, the first opening, and the second opening.

46. ​​The method according to any one of claims 40 to 44, wherein: The washing step includes forcing the wash buffer through the chromatographic membrane in a direction opposite to the first direction in which the liquid is supplied into the container in the separation step.

47. The method according to any one of claims 40 to 45, wherein: The eluting step includes supplying the eluting solution into the container and forcing the eluting solution to pass through the chromatographic membrane in a direction opposite to the first direction in which the liquid is supplied into the container in the separating step.

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