Dry fibrillated membranes and affinity chromatography devices comprising same

Abstract

The present disclosure relates to an affinity chromatography device comprising a dry fibrillated polymer film having a moisture content of less than or equal to 60 mass% of the dry fibrillated film. When the dry fibrillated polymer film is formed, the non-dry fibrillated polymer film may be dried by freeze-drying. The dry affinity chromatography device may improve the stability or viability of the target molecule being purified, reduce shipping risk (e.g., reduce the risk of accidental rupture or leakage due to falling), and reduce storage risk (e.g., reduce the risk of microbial growth during storage). The affinity chromatography device and / or the dry fibrillated polymer film may be treated with ethylene oxide, gamma irradiation, or X-ray irradiation. In some embodiments, the treatment sterilizes the chromatographic device or dry fibrillated polymer film. The dry affinity chromatography device may be wetted with a bioprocessing fluid prior to use.

Description

Technical Field

[0001] The present disclosure relates generally to affinity chromatography and, more particularly, to dry fibrillated membranes containing inorganic particles therein and affinity chromatography devices comprising dry fibrillated polymer membranes that allow for the separation of target molecules from aqueous mixtures. Background Art

[0002] Chromatographic methods are commonly used to separate and / or purify target molecules, such as proteins, nucleic acids, and polysaccharides, from mixtures. Affinity chromatography specifically involves passing the mixture through a matrix having a specific ligand (i.e., a specific binding partner) that binds to the target molecule. Once exposed to the ligand, the target molecule binds to the matrix and is thus retained from the mixture. Affinity chromatography offers certain advantages over other types of chromatography. For example, affinity chromatography provides a purification method that can separate a target protein from a mixture of a target protein and other biomolecules in a single step with high yield.

[0003] Despite the advantages of current affinity chromatography devices, there is a need in the art for chromatography devices that can be used with shorter residence times than conventional devices while providing the same or better binding capacity than current products, are reusable, ready-to-use and / or sterilizable, and have reduced shipping and / or storage risks. Summary of the Invention

[0004] According to one aspect ("Aspect 1"), a composite membrane for an affinity chromatography device comprises: a dry fibrillated polymer membrane having inorganic particles therein, wherein at least one of the inorganic particles and the dry fibrillated polymer membrane has an affinity ligand covalently bonded thereto, the affinity ligand reversibly binding to a target molecule, and wherein the moisture content of the dry fibrillated polymer membrane is less than or equal to 60% by mass of the dry fibrillated membrane.

[0005] According to another aspect (Aspect 2) of Aspect 1, the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

[0006] According to yet another aspect (Aspect 3) of Aspect 1 or Aspect 2, the inorganic particles have a spherical shape and a nominal particle size of about 5 microns to about 20 microns.

[0007] According to another aspect (Aspect 4) of Aspects 1 to 3, the inorganic particles have a particle size distribution of D90 / D10 of 3 or less.

[0008] According to yet another aspect (Aspect 5) of any one of Aspects 1 to 4, the dry fibrillated polymer film comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0009] According to a further aspect of any one of aspects 1 to 5 (Aspect 6), the affinity ligand is selected from the group consisting of: protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, oligonucleotides, and combinations thereof.

[0010] According to another further aspect (Aspect 7) of any one of Aspects 1 to 6, the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0011] According to a further aspect (Aspect 8) of any one of Aspects 1 to 7, the dry fibrillated polymer film has a wound configuration.

[0012] According to a further aspect (Aspect 9) of any one of Aspects 1 to 8, the dry fibrillated polymer film has a stacked configuration.

[0013] According to a further aspect (Aspect 10) of any one of Aspects 1 to 9, the dry fibrillated polymer film is treated with ethylene oxide, gamma irradiation, or X-ray irradiation for sterilization.

[0014] According to one aspect ("Aspect 11"), an affinity chromatography device comprises: a fluid inlet, a fluid outlet, the fluid outlet being fluidically connected to the fluid inlet, a dry fibrillated polymer membrane, the dry fibrillated polymer membrane being positioned between the fluid inlet and the fluid outlet, and containing at least one inorganic particle, wherein at least one of the following is satisfied: (1) before drying, the moisture content of the dry fibrillated polymer membrane is less than or equal to 60% by mass of the dry fibrillated polymer membrane, or (2) the moisture content of the affinity chromatography device is less than or equal to 60% by mass of the affinity chromatography device, and wherein at least one of the dry fibrillated polymer membrane and the inorganic particle has an affinity ligand covalently bonded thereto, and the affinity ligand reversibly binds to the target molecule.

[0015] According to a further aspect (Aspect 12) of Aspect 11, a housing is included that surrounds the fluid inlet, the fluid outlet, and the dry fibrillated polymer film.

[0016] According to yet another aspect (Aspect 13) of Aspect 11 or Aspect 12, two or more of the dry fibrillated membranes are in the form of a dry stacked membrane assembly positioned between the fluid inlet and the fluid outlet.

[0017] According to a further aspect (Aspect 14) of Aspect 11 or Aspect 12, the dry fibrillated film has a wound film construction comprising a dry fibrillated polymer wound around a core.

[0018] According to another further aspect of any one of aspects 11 to 14 (Aspect 15), the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0019] According to yet another aspect (Aspect 16) of any one of Aspects 11 to 15, the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

[0020] According to yet another aspect (Aspect 17) of any one of Aspects 11 to 16, the dry fibrillated polymer film comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0021] According to a further aspect of any one of aspects 11 to 17 (Aspect 18), the affinity ligand is selected from the group consisting of: protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, and combinations thereof.

[0022] According to a further aspect (Aspect 19) of any one of Aspects 11 to 18, at least one of the dry fibrillated polymer membrane and the affinity chromatography device is treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0023] According to a further aspect (Aspect 20) of any one of Aspects 11 to 19, the affinity chromatography device is configured to achieve a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds.

[0024] According to a further aspect of any one of aspects 11 to 20 (Aspect 21), the affinity chromatography device is configured to achieve about 100 (X 10 -12 cm 2 ) to about 2000(X 10 -12 cm 2 ) hydraulic permeability.

[0025] According to a further aspect of any one of aspects 11 to 21 (Aspect 22), the affinity chromatography device is configured to achieve an elution volume of 1 column volume (CV) to 6 column volumes (CV) from 100 mAU to 100 mAU.

[0026] According to a further aspect (Aspect 23) of any one of Aspects 11 to 22, the affinity chromatography device is configured to achieve a cycling durability of at least 100 cycles at an operating pressure of less than 0.3 MPa.

[0027] According to a further aspect (Aspect 24) of any one of Aspects 11 to 23, use of the article for separating a target molecule from a fluid stream.

[0028] According to another aspect (Aspect 25), the manifold comprises at least two affinity chromatography devices according to any one of Aspects 11 to 23 arranged in a parallel configuration.

[0029] According to another aspect (Aspect 26) of Aspect 25, the header is enclosed in a housing.

[0030] According to another aspect (Aspect 27), a device comprises a first header and a second header arranged in a parallel configuration, wherein each of the first header and the second header comprises at least two affinity chromatography devices according to any one of Aspects 11 to 23.

[0031] According to another aspect (Aspect 28) of Aspect 27, the first header and the second header are enclosed in a housing.

[0032] According to another aspect (Aspect 29), a method for separating target molecules includes: (1) providing a chromatographic device comprising an inlet, an outlet, and a dry fibrillated polymer membrane having a moisture content less than or equal to 60 mass% of the dry fibrillated membrane, wherein the dry fibrillated polymer membrane is positioned between the inlet and the outlet and contains at least one inorganic particle, (2) wetting the dry fibrillated polymer membrane to form a wet polymer membrane, (3) adding an aqueous mixture containing the target molecule to the inlet, (4) passing the aqueous mixture through the wet fibrillated polymer membrane, wherein the target molecule is bound to an affinity ligand, and (5) binding the target molecule to the affinity ligand, wherein at least one of the dry fibrillated polymer membrane and the inorganic particle has an affinity ligand thereon.

[0033] According to a further aspect (Aspect 30) of Aspect 29, comprising treating the dry fibrillated polymer film with ethylene oxide, thereby sterilizing the dry fibrillated polymer film.

[0034] According to a further aspect of aspect 30 (Aspect 31), the treatment is carried out according to the ethylene oxide (EO) treatment method of the membrane described in the Test Methods section.

[0035] According to yet another aspect (Aspect 32) of Aspect 29, the chromatographic device is treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0036] According to a further aspect of aspect 29 (Aspect 33), the method comprises treating the chromatography device with ethylene oxide, gamma irradiation, or X-ray irradiation to sterilize the chromatography device.

[0037] According to a further aspect of aspect 29 (aspect 34), comprising treating the non-dried fibrillated membrane so as to covalently bind the affinity ligand to one or both of the non-dried fibrillated polymer membrane and the inorganic particles.

[0038] According to a further aspect (Aspect 35) of Aspect 29, comprising characterizing the moisture content of the dry fibrillated polymer film.

[0039] According to a further aspect of aspect 35 (Aspect 36), the characterization is performed according to the method for determining the moisture content of the film described in the Test Methods section.

[0040] According to a further aspect of aspect 29 (Aspect 37), comprising characterizing the moisture content of the chromatography device.

[0041] According to a further aspect of aspect 29 (aspect 38), the method comprises drying the non-dried fibrillated film by freeze drying to form the dried fibrillated polymer film.

[0042] According to a further aspect of aspect 38 (Aspect 39), the drying is performed according to the freeze-drying method of the membrane described in the Test Methods section.

[0043] According to a further aspect (Aspect 40) of Aspect 29, the method comprises determining the dynamic binding capacity at 10% breakthrough.

[0044] According to a further aspect of aspect 40 (Aspect 41), the determining is performed according to the method for determining the dynamic binding capacity of a device article at 10% breakthrough as described in the Test Methods section.

[0045] According to a further aspect (Aspect 42) of Aspect 29, comprising initially wetting the dry chromatography device.

[0046] According to a further aspect of aspect 42 (Aspect 43), the initial wetting is performed according to the initial wetting method for dry device preparations described in the test methods section.

[0047] According to a further aspect (Aspect 44) of Aspect 29, the method comprises determining an elution volume of the chromatography device.

[0048] According to a further aspect of aspect 44 (Aspect 45), the determining is performed according to the method for determining the elution volume of the device preparation from 100 mAU to 100 mAU as described in the Test Methods section.

[0049] According to a further aspect (Aspect 46) of Aspect 29, comprising determining the dynamic binding capacity of the chromatography device at 10% breakthrough.

[0050] According to a further aspect of aspect 46 (Aspect 47), the determining is performed according to the method for determining the dynamic binding capacity of a device article at 10% breakthrough as described in the Test Methods section.

[0051] According to a further aspect of aspect 29 (Aspect 48), the chromatography device is an affinity chromatography device.

[0052] According to a further aspect of aspect 29 (Aspect 49), the affinity ligand is selected from the group consisting of: protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, and combinations thereof.

[0053] According to yet another aspect (Aspect 50) of Aspect 29, the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

[0054] According to yet another aspect (Aspect 51) of Aspect 29, the dry fibrillated polymer comprises: an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0055] According to another aspect (Aspect 52), the affinity chromatography device includes: a shell component, an inlet allowing fluid to flow into the shell component, an outlet allowing fluid to flow out of the shell component and connected to the inlet fluid, and a stacked membrane assembly, wherein the stacked membrane assembly is positioned within the shell component between the fluid inlet and the fluid outlet, wherein the stacked membrane includes two or more dry fibrillated polymer membranes in a stacked structure, each of the dry fibrillated polymer membranes containing inorganic particles therein, wherein the moisture content of the dry fibrillated polymer membrane is less than or equal to 60% by mass of the dry fibrillated polymer membrane, and at least one of the dry fibrillated polymer membrane and the inorganic particles has an affinity ligand covalently bonded thereto, and the affinity ligand reversibly binds to the target molecule.

[0056] According to another aspect (Aspect 53) of Aspect 52, the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0057] According to another aspect (Aspect 54) of Aspect 51 or Aspect 52, the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

[0058] According to yet another aspect (Aspect 55) of any one of Aspects 52 to 54, the inorganic particles have a spherical shape and a nominal particle size of about 5 microns to about 20 microns.

[0059] According to a further aspect (Aspect 56) of any one of Aspects 52 to 55, the particle size distribution has a D90 / D10 of less than or equal to 3.

[0060] According to yet another aspect (Aspect 57) of any one of Aspects 52 to 56, the dry fibrillated polymer film comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0061] According to a further aspect of any one of aspects 52 to 57 (Aspect 58), the affinity ligand is selected from the group consisting of: protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, oligonucleotides, and combinations thereof.

[0062] According to a further aspect of any one of aspects 52 to 58 (Aspect 59), the device is treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0063] According to another aspect (Aspect 60) of any one of Aspects 52 to 59, the dry fibrillated polymer film has a stacked configuration having two sides and edges of a plurality of stacked film assemblies, and wherein the device further comprises a material to seal the edges of the stacked film assemblies.

[0064] The foregoing embodiments are intended to be limiting and should not be construed as limiting or otherwise narrowing the scope of any inventive concepts otherwise provided herein. Although a number of embodiments are disclosed, other embodiments will be readily apparent to those skilled in the art from the following detailed description, which shows and describes illustrative embodiments. Accordingly, the drawings and detailed description are to be regarded as illustrative in nature and not restrictive. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] The accompanying drawings are included to provide a further understanding of the disclosure and are incorporated in and constitute a part of this specification. They illustrate embodiments of the disclosure and together with the description serve to explain the principles of the disclosure.

[0066] Figure 1 is an exploded view of a chromatography device according to at least one embodiment, the chromatography device comprising a wound film assembly including a dry fibrillated polymeric membrane having inorganic particles therein;

[0067] Figure 2 is a cross-sectional view of a chromatography device according to at least one embodiment, the chromatography device comprising a wound membrane assembly, the cross-sectional view depicting a dry fibrillated polymer membrane, an outer flow channel, and an inner flow channel;

[0068] Figure 3is an exploded view of a chromatography device according to at least one embodiment, the chromatography device comprising a stacked membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein;

[0069] Figure 4A is a cross-sectional view of a chromatography device according to at least one embodiment, the chromatography device comprising a stacked membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein;

[0070] Figure 4B According to at least one embodiment Figure 4A Exploded view of the mid-chromatographic device;

[0071] Figure 5 is a side view of a manifold having a plurality of chromatography devices in a parallel configuration, wherein each chromatography device contains a stacked membrane assembly comprising a dry fibrillated polymeric membrane having inorganic particles therein, according to at least one embodiment;

[0072] Figure 6 is a schematic side view of two headers in a parallel configuration according to at least one embodiment;

[0073] Figure 7 is a side view of a manifold having a plurality of chromatography devices in a parallel configuration, each chromatography device comprising a wound film assembly comprising a dry fibrillated polymeric membrane having inorganic particles therein, according to at least one embodiment;

[0074] Figure 8 is a side view of two headers in a side-by-side configuration containing wound dry fibrillated polymer membrane assemblies according to at least one embodiment.

[0075] Figure 9 is a chromatogram of UV absorption at 280 nm relevant to the characterization of device T according to at least one embodiment;

[0076] Figure 10A is a schematic diagram of a multiwell plate according to at least one embodiment; and

[0077] Figure 10B According to at least one embodiment Figure 10A Schematic diagram of a portion of a porous plate depicted in FIG, showing a portion of a stacked membrane assembly located on a porous substrate. DETAILED DESCRIPTION

[0078] Those skilled in the art will readily appreciate that the drawings referred to herein are not necessarily drawn to scale but may be exaggerated in order to illustrate aspects of the present disclosure, and therefore, the drawings should not be construed as limiting.

[0079] As used herein, the term "on" is intended to mean that an element (such as a polymer film) is directly on another element, or intervening elements may be present.

[0080] As used herein, the expressions "spherical particles," "spherical inorganic particles," and "inorganic particles having a spherical shape" may be used interchangeably.

[0081] As used herein, the term "fibrillated polymer film" is intended to mean that fibrils are present in the polymer film, e.g., the polymer film has a microstructure characterized by knots, fibrils, and voids, wherein the knots are interconnected by fibrils and the voids are the spaces between the knots and fibrils.

[0082] The terms "fibrillated polymer film," "polymer film," and "film" are used interchangeably herein.

[0083] As used herein, the term "spirally wound membrane assembly" is intended to include a dry fibrillated polymeric membrane alone, and a dry fibrillated polymeric membrane and one or more intermediate materials in a spiral configuration.

[0084] As used herein, the term "stacked film assembly" is intended to include dry fibrillated polymer films alone, and dry fibrillated polymer films and one or more intermediate materials in a stacked configuration.

[0085] As used herein, the terms "dry affinity chromatography device," "dry chromatography device," "dry device," and "dry device article" are used interchangeably.

[0086] The term "wet fibrillated polymer film" as used herein is intended to mean that a dry fibrillated film or a dry fibrillated film assembly has undergone a wetting process.

[0087] Herein, the terms "non-dry," "wet," and "moistened" are used interchangeably and are intended to refer to a dry fibrillated membrane or dry chromatography device prior to undergoing a drying process.

[0088] As used herein, the term "dry" may be defined as having a moisture content of less than or equal to 60% by mass of the dry fibrillated polymer film, or less than 55% by mass of the dry film, or less than 50% by mass of the dry film, or less than 45% by mass of the dry film, or less than 40% by mass of the dry film, or less than 35% by mass of the dry film, or less than 30% by mass of the dry film, or less than 25% by mass of the dry film, or less than 20% by mass of the dry film, or less than 15% by mass of the dry film, or less than 10% by mass of the dry film, or less than 5% by mass of the dry film, or less than 3% by mass of the dry film, or less than 1% by mass of the dry film, or less than 0.1% by mass of the dry film, or 0% by mass of the dry film.

[0089] As used herein, the term "dry" may also be defined as, or alternatively defined as, a moisture content of less than or equal to 60% by mass of the dry affinity chromatography device, or less than 55% by mass of the dry device, or less than 50% by mass of the dry device, or less than 45% by mass of the dry device, or less than 40% by mass of the dry device, or less than 35% by mass of the dry device, or less than 30% by mass of the dry device, or less than 25% by mass of the dry device, or less than 20% by mass of the dry device, or less than 15% by mass of the dry device, or less than 10% by mass of the dry device, or less than 5% by mass of the dry device, or less than 3% by mass of the dry device, or less than 1% by mass of the dry device, or less than 0.1% by mass of the dry device, or 0% by mass of the dry device.

[0090] As used herein, the term "about" means a measurement + / - 10% of the specified unit.

[0091] The present disclosure relates to dry fibrillated polymer membranes, whose moisture content is less than or equal to 60% by mass of the dry membrane. The dry fibrillated polymer membranes can be wound or stacked to be positioned in an affinity chromatography device. The moisture content of the affinity chromatography device (containing one or more dry fibrillated polymer membranes) can be less than or equal to 60% by mass of the device. In addition, the dry fibrillated polymer membrane contains inorganic particles (e.g., spherical or irregularly shaped inorganic particles) in and / or on the membrane. Affinity ligands can be combined with the inorganic particles and / or the dry fibrillated polymer membrane. When forming the dry fibrillated polymer membrane, non-dried fibrillated polymer membranes can be dried by freeze drying. The process of freeze drying simplifies the device manufacturing process, wherein before the fibrillated membrane is integrated into the chromatography device, the ligand chemical fixation to the membrane can be completed on the fibrillated membrane, thereby removing the in situ chemical fixation step from the device manufacturing process.

[0092] The affinity chromatography device and the dry fibrillated polymer membrane can be treated with ethylene oxide, gamma irradiation, X-ray irradiation, or any other treatment that sterilizes the device or membrane. In some embodiments, the dry fibrillated polymer membrane can be treated separately before assembling the chromatography device. In some embodiments, the dry fibrillated polymer membrane can be treated using the chromatography device after assembly.

[0093] The chromatography device (in any form, such as, but not limited to, an affinity chromatography device, a cartridge, a cartridge assembly, a manifold, a manifold chromatography device, a multi-port manifold, and / or a multi-port manifold chromatography device, as well as any other chromatography device described herein) may have a moisture content of less than or equal to 60% by mass of the dry affinity chromatography device, or less than about 55% by mass of the dry device, or less than about 50% by mass of the dry device, or less than about 45% by mass of the dry device, or less than about 40% by mass of the dry device, or less than about 35% by mass of the dry device, or less than about 50% by mass of the dry device. The dry device comprises less than about 30% by mass of the dry device, or less than about 25% by mass of the dry device, or less than about 20% by mass of the dry device, or less than about 15% by mass of the dry device, or less than about 10% by mass of the dry device, or less than about 5% by mass of the dry device, or less than about 3% by mass of the dry device, or less than about 1% by mass of the dry device, or less than about 0.1% by mass of the dry device, or less than about 0.01% by mass of the dry device, or less than about 0.001% by mass of the dry device, or 0% by mass of the device.

[0094] The dry fibrillated polymer film (in any form, such as, but not limited to, a fibrillated polymer film, a fibrillated film disc, a stacked film assembly, a wound film assembly, a spiral wound film assembly, a film assembly, and / or a fibrillated film layer, and any other fibrillated polymer film described herein) may have a moisture content of less than or equal to 60% by mass of the dry fibrillated polymer film, or less than about 55% by mass of the dry film, or less than about 50% by mass of the dry film, or less than about 45% by mass of the dry film, or less than about 40% by mass of the dry film, or less than about 10% by mass of the dry film. The present invention relates to an aqueous solution of at least about 1% by mass of the dry film, or less than about 35% by mass of the dry film, or less than about 30% by mass of the dry film, or less than about 25% by mass of the dry film, or less than about 20% by mass of the dry film, or less than about 15% by mass of the dry film, or less than about 10% by mass of the dry film, or less than about 5% by mass of the dry film, or less than about 3% by mass of the dry film, or less than about 1% by mass of the dry film, or less than about 0.1% by mass of the dry film, or less than about 0.01% by mass of the dry film, or less than about 0.001% by mass of the dry film, or 0% by mass of the dry film.

[0095] The dry fibrillated polymer film (in any form, such as, but not limited to, a fibrillated polymer film, a fibrillated film disc, a stacked film assembly, a wound film assembly, a spiral wound film assembly, a film assembly, and / or a fibrillated film layer, and any other fibrillated polymer film described herein) may have a moisture content of from about 0.001% to about 60% by mass of the dry fibrillated polymer film, from about 0.001% to about 55% by mass of the dry film, or from about 0.01% to about 50% by mass of the dry film. % by mass of the dry film, or about 0.05% to about 45% by mass of the dry film, or about 0.1% to about 40% by mass of the dry film, or about 0.5% to about 35% by mass of the dry film, or about 0.6% to about 30% by mass of the dry film, or about 0.7% to about 25% by mass of the dry film, or about 0.8% to about 20% by mass of the dry film, or about 0.9% to about 15% by mass of the dry film, or about 1% to about 10% by mass of the dry film. In some embodiments, the dry fibrillated polymer film has a moisture content of about 0.001% to about 5% by mass of the dry fibrillated polymer film, or about 0.01% to about 4.5% by mass of the dry film, or about 0.1% to about 4% by mass of the dry film, or about 1.0% to about 3.5% by mass of the dry film, or about 1.1% to about 3% by mass of the dry film. In some embodiments, the dry fibrillated polymer film has a moisture content of about 1.3% to about 2% by mass of the dry film.

[0096] The dry chromatography device (in any form, such as, but not limited to, an affinity chromatography device, a cartridge, a cartridge assembly, a manifold, a manifold chromatography device, a multi-port manifold, and / or a multi-port manifold chromatography device, as well as any other chromatography device described herein) may have a moisture content of about 0.001% to 60% by mass of the dry affinity chromatography device, about 0.001% to about 55% by mass of the dry device, or about 0.01% to about 50% by mass of the dry device, or about 0.01% to about 50% by mass of the dry device. In some embodiments, the chromatographic device has a moisture content of about 0.05% to about 45% by mass, or about 0.1% to about 40% by mass of the dry device, or about 0.5% to about 35% by mass of the dry device, or about 0.6% to about 30% by mass of the dry device, or about 0.7% to about 25% by mass of the dry device, or about 0.8% to about 20% by mass of the dry device, or about 0.9% to about 15% by mass of the dry device, or about 1% to about 10% by mass of the dry device. In some embodiments, the chromatographic device has a moisture content of about 0.001% to about 5% by mass of the dry chromatographic device, or about 0.01% to about 4.5% by mass of the dry device, or about 0.1% to about 4% by mass of the dry device, or about 1.0% to about 3.5% by mass of the dry device, or about 1.1% to about 3% by mass of the dry device. In some embodiments, the chromatographic device has a moisture content of about 1.3% to about 2% by mass of the dry device.

[0097] In some embodiments, the dynamic binding capacity (DBC) of the chromatography device at 10% penetration is greater than 35 mg / ml when the residence time is 20 seconds. In addition, the cyclic durability of the affinity chromatography device is at least 100 cycles at an operating pressure of no more than 0.3 MPa. In a further embodiment, the elution volume of the affinity chromatography device from 100 mAU to 100 mAU may be from about 1 to about 6 column volumes. In some embodiments, the dry affinity chromatography device can improve the stability or viability of the target molecule being purified. According to some embodiments, the dry affinity chromatography device can reduce storage risks because eliminating the fluid inside the device reduces the risk of microbial growth during storage. According to some embodiments, the dry affinity chromatography device can reduce shipping risks because eliminating the fluid inside the device reduces the risk of accidental rupture or leakage due to falling. In some embodiments, the affinity chromatography device containing a dry fibrillated polymer membrane can also shorten the lead time for shipping and distribution. After storage and / or shipment, the dry affinity chromatography device can then be readily wetted for use with bioprocessing fluids.

[0098] In some embodiments, dry affinity chromatography devices may have higher permeability and / or shelf life under ambient conditions. Affinity chromatography devices according to the present disclosure may have other benefits, such as: uniform flow fronts for vertical flows in a variety of configurations (e.g., stacked, spirally wound, cassettes, and parallel manifolds), enabling users to incorporate multiple dry affinity chromatography devices configured as parallel manifolds, and / or performance can be scaled by residence time. It will be understood that the terms "manifold" and "parallel manifolds" are used interchangeably herein and are intended to define the same configuration. Dry affinity chromatography devices containing dry fibrillated polymer membranes enable various processing modes for the final device (e.g., capable of being sterilized by ethylene oxide treatment, gamma treatment, X-ray treatment before or after assembly). This sterilization method is more limited or impossible when using existing wet product device formats.

[0099] In use, the dry affinity chromatography device is first wetted with a liquid suitable for bioprocessing, including but not limited to aqueous buffer solvents, harvests, acids, bases, organic solvents, and other fluids commonly used and known to those skilled in the art of bioprocessing. It will be understood that the chromatography device described herein is considered "dry" if it contains a dry fibrillated polymer membrane in any form.

[0100] Figure 1 and 21 and 2 are exploded views and cross-sectional views, respectively, of a chromatography device 100 according to at least one embodiment, comprising a wound film. In some embodiments, when forming the chromatography device 100, at least one inner intermediate material 200 may be placed around (e.g., wrapped around) a cylindrical core 150 to a desired width or a predetermined amount. Subsequently, a dry fibrillated polymer membrane containing spherical and / or non-spherical inorganic particles 210 is wrapped around the core 150 to cover the inner intermediate material 200 to a desired width or a predetermined amount. Subsequently, an outer layer of at least one outer intermediate material 220 is circumferentially placed on (e.g., wrapped around) the dry fibrillated polymer membrane 210 to a desired width or a predetermined amount. The combination of the inner intermediate material 200, the dry fibrillated polymer membrane 210, and the outer intermediate material 220 will be referred to herein as a "wound film assembly." In some embodiments, the "wrap film assembly" may also contain any combination of polymers and / or polymer intermediate materials wrapped around a cylindrical core. The cylindrical core 150 may have a hollow or solid interior. In either case, the core 150 has a solid outer wall, so that the aqueous mixture flowing through the chromatography device 100 flows within the internal flow channel 140 formed by the internal intermediate material 200, rather than flowing into the core 150. The use of a hollow core 150 reduces the amount of material used to form the core 150, reduces the weight of the device 100, and reduces manufacturing costs.

[0101] like Figure 1 As shown in FIG, the wrap assembly 110 includes an inner intermediate material 200, a dry fibrillated polymer film 210, and an outer intermediate material 220. The wrap assembly 110 and the central core 150 may be located within a housing 50. In some embodiments, the housing 50 is cylindrical. Figure 2 As shown in FIG, the outer intermediate material 220 forms the outer flow channel 130, while the inner intermediate material 200 forms the inner flow channel 140. It should be understood that the intermediate materials 200 and 220 in the embodiments described herein may be different or may be the same. In addition, two or more intermediate materials may be used to form one or both of the outer flow channel 130 and the inner flow channel 140. Non-limiting examples of suitable materials for forming the housing of the chromatography device described herein include, but are not limited to, polyurethane, stainless steel, polypropylene, acrylonitrile butadiene styrene (ABS), polyethylene terephthalate (PET), polyether ether ketone (PEEK), cyclic olefin copolymer (COC), and polyethylene terephthalate glycol (PETG). In addition, the shape of the housing unit is not limited and can take any form as long as the form encapsulates the affinity chromatography device and / or cartridge (described below) and / or manifold (described below).

[0102] When in use, the dry fibrillated polymer membrane 210 is first wetted as described above. After wetting, the aqueous mixture flows into the inlet 80 located in the inlet cover 60. The mixture flows over the distributor cover 65 and is directed to the outer flow channel 130 formed by the outer intermediate material 22. The distributor cover 65 directs the aqueous mixture to the outer flow channel 130 (i.e., the intermediate material 220) at approximately 90 degrees in the feed direction. This redirection promotes a more uniform flow of the aqueous mixture into the outer flow channel 130. The outer intermediate material 220 forming the outer flow channel 130 is located between the housing 50 and the wound dry fibrillated polymer membrane 210. The distributor cover 65 can be a polyolefin or coated with a polyolefin.

[0103] The aqueous mixture flows through the outer flow channel 130 (i.e., the outer intermediate material 220) and passes through the wound "wet" fibrillated polymer membrane 210 in a vertical direction (e.g., from the outer flow channel 130 to the inner flow channel 140). When the aqueous mixture passes through the wound wet fibrillated polymer membrane 210 in a vertical flow manner from the outer flow channel 130 (i.e., the outer intermediate material 220), the affinity ligand reversibly binds to the target molecule, thereby effectively removing it from the aqueous mixture. The aqueous mixture from which the target molecule has been removed then enters the inner flow channel 140 (i.e., the inner intermediate material 200) located between the solid outer wall of the central core 150 and the wound wet fibrillated polymer membrane 210. Subsequently, the aqueous mixture is redirected by the outlet cover 75 at the bottom of the inner flow channel 140. Subsequently, the aqueous mixture flows out of the chromatography device 100 through the outlet 85 located in the outlet cover 75.

[0104] The diameter and / or height of the central core 150, and / or the width and / or height of the dry fibrillated polymer membrane and / or intermediate material can be adjusted to achieve a larger volume without negatively impacting the performance of the chromatography device. In addition, the target molecule can be removed from the affinity ligand, for example, by passing a fluid having a lower pH through the chromatography device, as is known to those skilled in the art.

[0105] As long as the aqueous mixture can flow through, the intermediate materials 200 and 220 are not particularly limited. Some non-limiting examples of suitable intermediate materials include, but are not limited to: porous fluoropolymer membranes or porous non-fluoropolymer membranes (e.g., porous polypropylene or other porous polyolefin membranes), porous nonwoven materials, or porous woven materials. In some embodiments, the intermediate membranous object is a thermoplastic or thermosetting polymer membranous object. In some embodiments, the winding film assembly includes an integrated inlet end cap 60 and / or an integrated outlet end cap 75 located at opposite ends of the housing 50, thereby forming an integrated, reusable chromatographic device. In some embodiments, the inlet or inlet end cap 60 and the outlet or outlet end cap 75 may be located on the same side of the dry chromatography device 100.

[0106] In some embodiments, the affinity chromatography device 100 or the dry fibrillated polymer membrane 210 can be treated with ethylene oxide, gamma irradiation, X-ray irradiation, or any other sterilization / treatment method that sterilizes the device 100 or membrane 210. In some embodiments, the dry fibrillated polymer membrane 210 can be sterilized separately prior to assembly of the affinity chromatography device 100. In some embodiments, the dry fibrillated polymer membrane 210 can be sterilized after assembly of the affinity chromatography device 100.

[0107] In some embodiments, for example, Figure 3 In the embodiment generally depicted in the figure, the chromatography device 300 contains a dry fibrillated polymer membrane configured as dry fibrillated polymer membrane discs 340 stacked on top of each other to form a stacked membrane assembly 320. The dry fibrillated polymer membrane discs 340 can be placed in a stacked configuration simply by superimposing the dry fibrillated polymer membrane discs 340 on each other. Alternatively, the dry fibrillated polymer membrane discs 340 can be stacked and then laminated together by heat and / or pressure, or any other conventional method. The stacked membrane assembly 320 described herein is with respect to dry fibrillated polymer membrane discs for the sake of illustration. The dry fibrillated polymer membrane can be formed into other geometric shapes and / or non-geometric formations and is considered to be within the scope of the present disclosure. It should be understood that if the affinity chromatography device 300 contains a dry fibrillated polymer membrane or dry fibrillated polymer membrane discs 340, then the chromatography device 300 is considered to be "dry".

[0108] continue Figure 3 , the affinity chromatography device 300 includes: at least one upper intermediate material 360 located on the inlet side of the stacked membrane assembly 320, and at least one lower intermediate material 380 located on the outlet side of the stacked membrane assembly 320. The upper and lower intermediate materials 360, 380 may be the same or different, respectively. Similar to the wound film assembly described above, as long as the aqueous mixture can flow through, the intermediate materials 360, 380 used to form the stacked membrane assembly 320 are not particularly limited. Non-limiting examples of suitable intermediate materials include, but are not limited to: porous fluoropolymer membranes or porous non-fluoropolymer membranes (e.g., porous polypropylene or other porous polyolefin membranes), porous nonwoven materials, or porous woven materials.

[0109] The stacked membrane assembly 320 can be disposed in a housing 350 having an inlet cap 365 and an outlet cap 375 disposed at opposite ends of the housing 350. In some embodiments, for example, wherein the dry fibrillated polymer membrane is disc-shaped or circular, the housing 350 is cylindrical, although any geometric shape that is capable of wrapping the stacked membrane assembly and achieving the desired dynamic binding capacity is within the scope of the present disclosure. In some embodiments, the intermediate materials 360, 380, the housing 350, the inlet cap 365, and the outlet cap 375 can be formed from a thermoplastic polymer (such as polypropylene, polyethylene, or other polyolefins). Alternatively, one or both of the intermediate materials 360, 380 can be formed from an inorganic or metallic material, as long as the porous intermediate materials 360, 380 do not interfere with the operation of the chromatography device.

[0110] The dry fibrillated polymer membrane disc 340 in the stacked membrane assembly 320 can be adhered to the housing 350 at the inner wall of the housing 350 by any conventional method that prevents flow between the dry fibrillated polymer membrane disc 340 and the housing 350 (e.g., melt sealing or the use of a sealant). The inlet cover 365 and the outlet cover 375 can be sealed to the housing 350 by a similar or identical process. The inlet cover 365 and the outlet cover 375 include an inlet 380 and an outlet 385, respectively, to allow the aqueous mixture to flow into and out of the affinity chromatography device 300. Specifically, the inlet 380 allows fluid to flow into the housing 350, while the outlet 385 allows fluid to flow out of the housing 350. Before the chromatography device 300 is used, the dry fibrillated polymer membrane disc 340 is kept dry. Prior to use, the dry affinity chromatography device 300 or the dry fibrillated polymer membrane / dry fibrillated membrane disk 340 is wetted with a fluid suitable for bioprocessing, including but not limited to: aqueous buffer solutions, harvested materials, acids, bases, organic solvents, or other fluids commonly used and known to those skilled in the art of bioprocessing.

[0111] In use, the aqueous mixture flows sequentially through the upper intermediate material 360, through the "wetted" fibrillated polymer membrane disc 340 forming the stacked membrane assembly 320, and the lower intermediate material 380. As the aqueous mixture flows through the "wetted" fibrillated polymer membrane disc 340, the affinity ligand reversibly binds to the target molecule, effectively removing it from the aqueous mixture. The target molecule can be removed from the affinity ligand, for example, by passing a fluid with a lower pH value through the device, as is known to those skilled in the art.

[0112] In some embodiments, the affinity chromatography device 300 and / or the dry fibrillated polymer membrane disk 340 can be treated with ethylene oxide, gamma irradiation, X-ray irradiation, or other treatment methods that can sterilize the device 300, including sterilizing the dry fibrillated membrane disk 340. In some embodiments, the stacked membrane assembly 320 or the dry fibrillated polymer disk can be sterilized using any of the aforementioned sterilization methods prior to assembling the affinity chromatography device.

[0113] The total number of dry fibrillated polymer films present in the stacked film assembly is not particularly limited, but depends on the desired end use and / or the desired mass transport flow within the stacked film assembly. The stacked film assembly may include a total of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 (or more) dry fibrillated polymer films. It should be understood that there may be hundreds or even thousands of dry fibrillated polymer films in the stacked film assembly.

[0114] As previously described, the dry fibrillated polymer film contains inorganic particles therein. In some embodiments, the dry fibrillated polymer film may contain one or more types of inorganic particles, and / or may contain one or more nominal particle sizes within the dry fibrillated polymer film. Non-limiting examples of suitable inorganic particles include, but are not limited to, silica, zeolites, hydroxyapatite, metal oxides, and combinations thereof. It should be understood that the term "silica" as used herein is intended to describe silica that does not contain any measurable amount of boron, or does not contain boron by X-ray photoelectron spectroscopy (XPS). In addition, the inorganic particles may be solid or porous, and may have various sizes and shapes.

[0115] The inorganic particles may have a nominal particle size of about 0.1 micron, about 0.5 micron, about 1 micron, about 5 microns, about 10 microns, about 15 microns, about 20 microns, or about 25 microns, or larger. In addition, the inorganic particles may be monodisperse or polydisperse.

[0116] The dry fibrillated polymer film in both the winding film assembly and the stacked film assembly may contain spherical inorganic particles therein, or particles with a spherical configuration. As used herein, the term "spherical" refers to that the inorganic particles have a circular or nearly circular shape, wherein the distance from the center of the inorganic particle to any point on the outer edge of the particle is the same or nearly the same distance. In some embodiments, the spherical inorganic particles have a particle size distribution wherein D90 / D10 is less than or equal to 3, less than or equal to 2.5, less than or equal to 2, less than or equal to 1.5, or less than or equal to 1. The nominal particle size of the spherical inorganic particles may be about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations and blends thereof. In some embodiments, the spherical inorganic particles are polydisperse.

[0117] In some embodiments, the affinity ligand is covalently bonded to the inorganic particles. In another embodiment, the affinity ligand is covalently bonded to the dry fibrillated polymer film. In a further embodiment, the affinity ligand may be bound to both the dry fibrillated polymer film and the inorganic particles. The affinity ligand may be a protein, an antibody, or a polysaccharide that reversibly binds to a target protein or antibody. In one embodiment, the affinity ligand is a protein that reversibly binds to, for example, the Fc region of an antibody, an antibody fragment, an Fc fusion protein, or an antibody / drug conjugate. In another embodiment, the affinity ligand is an antibody, protein L, or a polysaccharide that reversibly binds to a specific protein or protein fragment. The affinity ligands used in the affinity chromatography device may include, but are not limited to, protein A, protein G, protein L, human Fc receptor protein, antibodies that specifically bind to other proteins, and heparin. The affinity ligand may be natural, recombinant, or synthetic. In some embodiments, the affinity ligand may be an oligosaccharide or oligonucleotide that reversibly binds to a nucleic acid (including but not limited to mRNA). In some embodiments, the affinity ligand may be an antibody or polysaccharide that reversibly binds to a viral vector (including but not limited to adeno-associated virus). In another embodiment, the affinity ligand is a metal affinity ligand that reversibly binds to a His-tag labeled protein.

[0118] According to some embodiments, the dry fibrillated polymer film comprises: expanded polytetrafluoroethylene film, expanded modified polytetrafluoroethylene film, expanded tetrafluoroethylene copolymer film, or expanded polyethylene film. In some embodiments, the dry fibrillated polymer film comprises expanded polytetrafluoroethylene film or expanded polyethylene film.

[0119] Although embodiments of the wound film assembly 110 and the stacked film assembly 320 are described herein, it should be understood that any number of dry fibrillated polymer films and any and all combinations of dry fibrillated polymer film types, spherical inorganic particle types, spherical inorganic particle sizes, and orientations of the dry fibrillated polymer films within the film assemblies 110, 320 are within the scope of the present disclosure. In addition, some or all of the dry fibrillated polymer films may differ from one another in composition, thickness, permeability, etc.

[0120] Figure 4A is a cross-sectional view of a chromatography device 400 including a cassette assembly 401; Figure 4B yes Figure 4A : an exploded view of the chromatography device 400 shown in . The cassette assembly 401 includes a dry fibrillated polymer membrane assembly containing inorganic particles therein. The dry fibrillated polymer membrane assembly can be configured as dry fibrillated polymer membrane layers 440 stacked on top of each other or parallel to each other, thereby forming a stacked membrane assembly 410. In some embodiments, the dry fibrillated polymer membrane layers 440 can be positioned in a stacked configuration by simply placing the dry fibrillated polymer membrane layers 440 adjacent to each other. Alternatively, the dry fibrillated polymer membrane layers 440 can be stacked and subsequently stacked together (e.g., spaced apart throughout the membrane layer, or connected at the edges of each membrane layer). As shown, the affinity chromatography device 400 (e.g., cassette assembly 401) includes a fluid inlet flow channel 402 connected to an inlet housing 404, and a fluid outlet flow channel 406 connected to an outlet housing 408. The fluid inlet flow channel 402 is fluidically connected to the fluid outlet flow channel 406. In some embodiments, device 400 includes a dry fibrillated polymeric membrane assembly 410 having at least one inorganic particle contained therein, positioned between inlet housing 404 and outlet housing 408. It will be understood that a chromatography device 400 is considered "dry" if it contains a dry fibrillated polymeric membrane.

[0121] Chromatography device 400 optionally includes: at least one inlet side intermediate material 460, which is positioned at the inlet side of dry stacked membrane assembly 410, and at least one outlet side intermediate material 480, which is positioned at the outlet side of dry stacked membrane assembly 410. The inlet side and outlet side intermediate materials 460, 480 can be the same or different. Similar to the wound and stacked membrane assemblies described above, the inlet side and outlet side intermediate materials 460, 480 used to form the cassette stacked membrane assembly 410 are not particularly limited, as long as the aqueous mixture can flow through them. Non-limiting examples of suitable intermediate materials include, but are not limited to: porous fluoropolymer membranes or porous non-fluoropolymer membranes (e.g., porous polypropylene or other porous polyolefin membranes), porous nonwoven materials, or porous woven materials. In some embodiments (not shown), there are no inlet side and outlet side intermediate materials 460, 480.

[0122] In some embodiments, at least one of the dry fibrillated polymer membrane layer 440 and the inorganic particles has an affinity ligand covalently bonded thereto, wherein the affinity ligand reversibly binds to the target molecule. In some embodiments, the affinity chromatography device 400 can be treated with ethylene oxide, gamma irradiation, X-ray irradiation, or other treatment methods known to those skilled in the art to sterilize the dry fibrillated polymer membrane layer 440 or the chromatography device 400.

[0123] like Figure 4A and 4B As shown in FIG, the inlet housing 404 and the outlet housing 408 are substantially planar and are stacked on opposite sides of the dry stacked film assembly 410 or parallel to opposite sides of the dry stacked film assembly 410 to form a box assembly 401. The dry stacked film assembly 410 contains a dry fibrillated polymer film layer 440. In some embodiments, the inlet housing 404 and the outlet housing 408 have substantially the same shape as the dry fibrillated polymer film assembly 410. For example, Figure 4A and Figure 4B As shown in FIG, both housings 404, 408 and dry stack film assembly 410 have a substantially rectangular shape. In some embodiments, housings 404, 408 and dry stack film assembly 410 can be any shape as long as housings 404 and 408 completely encompass dry stack film assembly 410.

[0124] In some embodiments, the dry stacked film assembly 440 (and the dry fibrillated polymer film layer 440) may include a perimeter sealing material 412 to seal the edges of the stacked film assembly 410. The perimeter sealing material 412 is not particularly limited, as long as the aqueous mixture can flow through the stacked film assembly 410. Some non-limiting examples of suitable perimeter sealing materials include, but are not limited to, polyolefins, thermoplastic elastomers, or combinations thereof.

[0125] For example, in some embodiments, and as Figure 4B , the inlet-side and outlet-side intermediate materials 460, 480 may be external to the perimeter sealing material 412. In some embodiments, the inlet-side and outlet-side intermediate materials 460, 480 may each comprise a separate intermediate perimeter sealing material that is distinct from the perimeter sealing material 412 that seals the edges of the stacked membrane assembly 410. In still other embodiments (not shown), the inlet-side and outlet-side intermediate materials 460, 480 may be internal to, and thus sealed within, the perimeter sealing material 412.

[0126] Figure 55 is a side view of a parallel manifold chromatography device 500 having a plurality of chromatography devices (each of which is a cassette assembly), wherein each chromatography device comprises a dry fibrillated polymer membrane having inorganic particles therein. In some embodiments, the plurality of chromatography devices may comprise a mixture of "wet" and dry fibrillated polymer membranes.

[0127] As shown, the manifold chromatography device 500 includes six cassette assemblies 501a-f (eg, Figure 4A In some embodiments, the manifold may include anywhere from 2 to 20 parallel cassette assemblies. In some embodiments, the manifold may include more than 20 parallel cassette assemblies, as long as similar flow and permeability profiles are present throughout the cassette assembly. This similarity across the cassette assembly allows for scalability of device size and performance.

[0128] In some embodiments, the fluid inlet flow channel 502 may include an optional flow distributor 520 disposed upstream of the first device 501a in the plurality of devices 501a-f. The optional flow distributor 520 makes the flow more uniform and helps promote flow distribution through the fluid inlet flow channel 502.

[0129] In some embodiments, the manifold chromatography device 500 may include an optional gas separation device 530 (e.g., an integrated gas trap) upstream of the fluid inlet flow channel 502. It will be appreciated that the fluid inlet flow channel 502 and the fluid outlet flow channel 504 may be located on opposite sides of the manifold in the chromatography device, on the same side, or in other configurations, so long as fluid flows through each chromatography device in a parallel manner.

[0130] Figure 6 is a cross-sectional view of a multi-port header 650 having at least two headers 600A and 600B (e.g., Figure 5 The manifolds in the chromatography device 500 are positioned within the optional housing 620. Each manifold contains a cassette assembly comprising a dry fibrillated polymer membrane having inorganic particles therein.

[0131] like Figure 6 As shown in FIG, the multi-port header 650 includes two headers 600A and 600B (e.g. Figure 500B) are arranged in a parallel configuration. In some embodiments, the multi-port manifold may include between 2 and 20 manifolds parallel to each other. In some embodiments (not shown), the multi-port manifold may include more than 20 manifolds parallel to each other. It will be understood that the fluid inlet flow channel 652 and the fluid outlet flow channel 654 may be located on opposite sides of the manifold in the chromatographic device, on the same side, or in other configurations, as long as the fluid flows through each manifold 600A and 600B in a parallel manner. In some embodiments, the manifolds 600A, 600B may each include a fluid outlet flow channel 654A, 654B, and an optional gas separation device 630A, 630B (e.g., an integrated gas trap) located upstream of the fluid inlet flow channels 652A, 652B. In a further embodiment (not shown), an optional gas separation device may be provided upstream of the fluid inlet flow channel 652 of the multi-port manifold device.

[0132] Figure 7 7 is a side cross-sectional view of a manifold 750 having a plurality of chromatography devices comprising wound film assemblies. Each wound film assembly comprises a dry fibrillated polymer membrane having inorganic particles therein. The chromatography devices are arranged in a parallel configuration. Figure 7 The depicted chromatography device is considered a dry chromatography device if it comprises a dry fibrillated polymer membrane or a dry wound film assembly.

[0133] In some embodiments, the affinity chromatography devices described herein can be used in a manifold 750, wherein the wound membrane affinity chromatography devices 700, 701 (e.g. Figure 1 and 2 The chromatographic device 100 described in Figure 7 In some embodiments, the manifold 750 may include an optional gas separation device 730 (eg, an integrated gas trap) upstream of the fluid inlet 740 . In some embodiments, the manifold 750 is located within the housing 720 .

[0134] In use, first use the fluid that is applicable to bioprocessing by the dry stacking membrane assembly or dry fibrillated polymer film wetting in each chromatographic device, described fluid includes but is not limited to: aqueous buffer solution, harvested material, acid, alkali, organic solvent and other fluid commonly used and known by those skilled in the art of bioprocessing. Subsequently, aqueous mixture flows into fluid inlet 740. Fluid inlet 740 separates the stream of aqueous mixture and enters at least two inlet pipes 760, 761. The split aqueous mixture in inlet pipes 760, 761 flows into chromatographic device 700, 701 subsequently, and wherein target molecule is captured by the affinity ligand on fibrillated polymer film and / or inorganic particles. Aqueous solution (it is the aqueous mixture removing the target molecule captured by affinity ligand) flows out from chromatographic device 700, 701 by outlet pipe 780, 781 respectively. The aqueous solution in outlet pipe 780, 781 merges in distribution element 790, and is reorganized into single aqueous solution.

[0135] Will understand, Figure 7 While two chromatography devices 700 and 701 are shown for illustrative purposes only, multiple, or greater, chromatography devices described herein can be used in a parallel configuration within a manifold, as long as similar flow and permeability profiles exist throughout the chromatography devices. This similarity between affinity chromatography devices allows for scalability of device size and performance. Furthermore, affinity chromatography devices can be incorporated into parallel configuration systems without requiring any changes or additions to the manifold 750. The fluid inlet 740 and fluid outlet 790 can be located on opposite sides of the manifold of the chromatography device, on the same side, or in other configurations, as long as fluid flow through each chromatography device 700 and 701 occurs in parallel.

[0136] Figure 8 is a cross-sectional view of two headers in a parallel configuration containing wound dry fibrillated polymer membrane assemblies according to at least one embodiment. Figure 8 As shown in FIG, the multi-port header 26 contains two headers 6a, 6b in a parallel configuration. The headers 6a and 6b (e.g. Figure 7 750 in the multi-port manifold 26) each contains at least two affinity chromatography devices 26a, 26b, and 26c, 26d. The ability to utilize at least two parallel manifolds advantageously enables increased volumetric capacity while utilizing the chromatography devices described herein. In some embodiments, the multi-port manifold 26 may include an optional gas separation device 830 (e.g., an integrated gas trap) upstream of the fluid inlet 30.

[0137] refer to Figure 10A and 10BAs described above, the dry fibrillated membrane in the form of a stacked membrane assembly can be fixed to a porous plate 1000, which contains a porous surface 1030 that separates a lower chamber (which can be operated at reduced pressure) from an upper chamber (operated at a higher pressure, such as atmospheric pressure). Figure 10A In the embodiment depicted in , a stacked film assembly formed of dry fibrillated polymer films 1020, each containing inorganic particles, may be used.

[0138] Test Method

[0139] Methods for determining membrane moisture content

[0140] The film moisture content was determined following the guidelines defined by ASTM Standard E1868-10 Standard Test Methods for Loss-On-Drying for Thermogravimetry. A Mettler Toledo HB43 moisture analyzer (Mettler-Toledo GmbH, CH-8606 Lake Griffin, Switzerland) was used for all data collection. A 109.5 cm 2 A constant film area was obtained by punching six 2 cm diameter circles. For wet samples, each circle was briefly placed on a low-lint wipe for a few seconds to remove surface moisture before loading into the instrument for measurement. Each sample was heated at 110°C for a total of 2 hours. Values ​​are reported as loss on drying (LOD) in mass %.

[0141] Methods for measuring membrane air permeability, thickness and density

[0142] To determine the density and thickness of each film sample, a 1 inch diameter (5.3 cm) 2 ) circles. The samples were weighed, and the thickness was measured using a Mitutoyo Litematic VL-50 thickness gauge (Mitutoyo America Corporation, Aurora, IL). The air permeability of the membranes was characterized using a Gurley Model 4340 Automatic Densitometer (Gurley Precision Instruments, 514 Fulton Street, Troy, NY 12180). Density was calculated by weighing each die-cut circle and dividing by the sample volume, which was calculated from the die-cut diameter and the measured thickness.

[0143] Methods for determining the dynamic binding and elution behavior of membranes

[0144] To determine the dynamic binding capacity of each membrane sample, two 25 mm diameter (4.9 cm) 2 These samples were stacked and held in a filter clamp, allowing controlled solution flow through the membrane.

[0145] Two 25 mm film sample discs were placed on the support screen ( Support Screen, stainless steel, 89428-948) and glass funnel ( Glass Funnel, 15ml, 89428-938), placed in a 125ml graduated filter bottle assembly ( Filter Flask Graduated,125ml,89428-978).

[0146] Place the sample bottle ( Sample bottle, clear borosilicate bottle, threaded, 20ml, 66009-567) into a 125ml filter bottle ( Graduated filter bottle, 125ml, 89428-978), where the outlet of the glass base ( The glass base of the stainless steel support, 89428-942) is located inside the sample bottle. The supporting screen, stainless steel, 89428-948) is placed on a glass base. The membrane sample is placed on the supporting screen and the glass funnel ( Glass funnel, 15ml, 89428-938) and by an aluminum fixture ( Aluminum clamp, 89428-944) for holding.

[0147] Using the following protocol, single binding and elution cycles were performed to determine the dynamic binding and elution properties of the membrane. The solutions described in Table A were applied to a glass funnel in the volumes and process order shown in Table B. The eluate from each process step was collected in a dedicated sample vial ( Sample bottles, clear borosilicate glass, threaded, 20 ml, 66009-567) were as shown in Table B. After each method step, the eluate sample bottle was replaced. Each solution flowed through the membrane under gravity.

[0148] Table A

[0149]

[0150] Table B

[0151]

[0152] The concentration of mAb in each elution sample was calculated by measuring the absorbance at a wavelength of 280 nm using a Hitachi U-2900 spectrophotometer (Hitachi High-Tech in America, 10 North Martingale Road, Suite 500, Schaumburg, Illinois 60173-2295).

[0153] The concentration of mAb in the target analyte feed and flowthrough and wash samples was calculated by measuring absorbance at 280 nm using a Hitachi U-2900 spectrophotometer (Hitachi High-Tech in America, 10 North Martingale Road, Suite 500, Schaumburg, Illinois 60173-2295).

[0154] The mAb concentration of the samples was calculated using Beer's law (Equation 01).

[0155] Formula -01A=e×L×C

[0156] in:

[0157] A = absorbance

[0158] e=1.47mL g -1 cm -1 (Extinction coefficient of target analyte, trastuzumab biosimilar)

[0159] L = sample path length

[0160] C = solution concentration

[0161] Membrane freeze-drying method

[0162] The membrane of albumin A is fixed in order to dry, these membranes are suspended and fully rinsed with deionized (DI) water, it is ensured that most of the solvent to be evaporated is water.Complete the final immersion in water, subsequently in Revco DXF-40040A ultra-low temperature freezer (Thermo Fisher Scientific, 81Wyman Street, Waltham, MA 02451) at -40 ℃ by each membrane freezing 24 hours.The frozen membrane of each suspension is transferred to VirTis 25LGenesis SQ Super ES 55 refrigerator (SPIndustries, 935Mearns Road, Warminster, PA 18974), wherein before starting circulation, condenser and rack (shelf) are both frozen, follow the step listed in Table C subsequently, under initial rack (≤-35 ℃), by membrane drying, realize vacuum (≤100mF) condition according to the program setting of circulation.

[0163] Table C

[0164]

[0165]

[0166] After completion of the freeze-drying cycle, the moisture content of each dried membrane was characterized according to the method for determining membrane moisture content.

[0167] Method for treating membrane with ethylene oxide (EO)

[0168] After drying following the lyophilization method for the membranes, each dried membrane subjected to ethylene oxide (EO) treatment was cut into pieces and placed into sterilization bags (Part No. PG-7514, plastcareusa, Canoga Park, CA).

[0169] Using a 3M Steri-Vac TM Some membranes were EO treated using a GS8X-1D sterilizer (3M Health Care, St. Paul, MN) where the treatment conditions represent an EO sterilization cycle. This EO treatment cycle used Steri-Gas TM An EO gas cartridge (Part No. 8-170, 3M Health Care, St. Paul, MN) injected 170 grams of EO into the sterilizer containing the membrane, and the sterilizer was maintained at a temperature of 40° C. for 3 hours. Details of this cycle can be found in Table D.

[0170] Table D

[0171]

[0172] Method for determining the dynamic binding capacity of device articles at 10% breakthrough

[0173] Insert the chromatography device into the AKTA TM A single cycle consisting of the following protocol was performed in the flow path of a Pure (Cytiva, Marlborough, MA) liquid chromatography system. Table E describes the solutions used, while Table F describes the protocol steps for determining the dynamic binding capacity at 10% breakthrough.

[0174] Table E

[0175] solution describe A 50 mM sodium phosphate, supplemented with 150 mM sodium chloride, pH ~ 7.4 B 100 mM citrate, pH ~3.4 CIP* 0.1M NaOH Target analytes 2.9-3.0 mg / ml polyclonal IgG (Lee Biosciences) dissolved in Solution A Preservative solution 20 / 80 v / v ethanol / water

[0176] * Clean in place

[0177] Table F

[0178]

[0179] Method for determining liquid permeability of a device article

[0180] The liquid permeability of the chromatographic device was determined using Darcy's law. The bed cross-sectional area and bed length of each device were characterized. Solution A was used as the fluid and its viscosity was characterized. TM The pressure drop across the column was measured as a function of liquid flux on a Pure Liquid Chromatography System (Cytiva, Marlborough, MA).

[0181] Method for drying initial wetness of device products

[0182] The dry device articles were wetted prior to characterization testing. The chromatography device was inserted into the flow path of an AKTA Pure liquid chromatography system and 50 mM sodium phosphate, 150 mM sodium chloride pH 7.4 buffer was fed through the device first in an upflow direction and then in a downflow direction, following the flow rates and volumes in Table G below.

[0183] Table G

[0184]

[0185] Method for determining the moisture content of device products

[0186] After assembly, the dry device articles were weighed to obtain the device dry mass. Subsequently, after initial wetting, the device articles were weighed again and permeability and DBC data were collected. The device wet mass represents the device when filled with the storage solution described in Table E.

[0187] Method for treating device products with ethylene oxide (EO)

[0188] Each dry device preparation that had undergone ethylene oxide (EO) sterilization was placed in a sterilization bag (Part No. PG-7514, plastcareusa, Canoga Park, CA).

[0189] Using a 3M Steri-Vac TM Some devices were EO treated using a GS8X-1D sterilizer (3M Health Care, St. Paul, MN) where the treatment conditions represent an EO sterilization cycle. This EO treatment cycle used Steri-Gas TM An EO gas cartridge (Part No. 8-170, 3M Health Care, St. Paul, MN) injected 170 grams of EO into the sterilizer containing the device, and the sterilizer was maintained at a temperature of 40° C. for 3 hours. Details of this cycle can be found in Table H.

[0190] Table H

[0191]

[0192] Method for determining the elution volume of a device preparation from 100 mAU to 100 mAU

[0193] Insert the wetted chromatography device into the AKTA TM A single cycle consisting of the following protocol was performed in the flow path of a Pilot 600 (Cytiva, Marlborough, MA) liquid chromatography system. Table I describes the solutions used, and Table J describes the purification protocol steps used. The elution volume from 100 mAU to 100 mAU was determined using the resulting chromatogram of UV absorbance at 280 nm, which was generated by following the purification protocol in Table J.

[0194] Table I

[0195]

[0196]

[0197] * Clean in place

[0198] Table J

[0199]

[0200] The resulting chromatogram of UV absorbance at 280 nm was plotted against the cumulative volume flowing through the device, generated by following the purification protocol in Table J, and used to determine the elution volume of the device preparation from 100 mAU to 100 mAU using Eq. 02.

[0201] Type-02EV 100至100 =CV100结束 -CV 100起始

[0202] in:

[0203] EV 100至100 = elution volume from 100 mAU to 100 mAU

[0204] CV 100起始 = the cumulative volume of buffer solution and CHO cell harvest (in column volumes, CV) that flowed through the device until the start of elution, which is defined as the point at which the UV absorbance at 280 nm rises above 100 mAU when the device is fed with elution buffer according to Table J, Step 4, and

[0205] CV 100结束 = the cumulative volume of buffer solution and CHO cell harvest (in column volumes, CV) that flowed through the device until the end of elution, which is defined as the point at which the UV absorbance at 280 nm drops below 100 mAU when the device is fed with elution buffer according to Table J, Step 4.

[0206] Example

[0207] Membrane A is a porous polytetrafluoroethylene (ePTFE) membrane having 15% by mass ePTFE and 85% by mass porous spherical silica particles. Membrane B is a porous ePTFE membrane having 15% by mass PTFE and 85% by mass porous irregularly shaped silica particles. Both membranes A and B are identical to those described in U.S. Patent Publication No. 2023 / 0356109 (attributed to Clinger et al.). Table K lists some physical properties of these two porous ePTFE membranes.

[0208] Table K

[0209]

[0210] The membrane was then treated in a manner that resulted in covalent bonding (immobilization) of Protein A to the ePTFE membrane. This method is typical of methods used by those skilled in the art and is further described in U.S. Pat. No. 10,525,376 (attributed to McManaway et al.) and U.S. Pat. No. 10,526,367 (attributed to McManaway et al.), resulting in Membranes C and D, respectively, to which Protein A was immobilized.

[0211] The protein A-immobilized membranes were dried via lyophilization as described in the above-mentioned method for lyophilizing membranes to obtain dried protein A-immobilized membranes E and F, respectively.

[0212] Cut pieces of the dried immobilized membranes E and F were subjected to ethylene oxide treatment as described above in the method for ethylene oxide (EO) treatment of membranes to yield dried protein A immobilized ethylene oxide treated membranes G and H, respectively.

[0213] The membranes made as described above were tested to evaluate their moisture content, density, thickness, and dynamic binding and elution behavior using the protocols described in the test methods described herein. Air permeability was also measured for each dried membrane. The performance of each of these affinity chromatography membranes is shown in Table L.

[0214] Table L

[0215]

[0216] Non-dried reference affinity chromatography devices in stacked membrane and parallel multi-port formats were made using non-dried membranes C and D (which were manufactured as described above). Dry affinity chromatography devices in stacked membrane, spiral wound, and parallel multi-port formats were made using dried membranes E to H (which were manufactured as described above). The membranes and conditions associated with each non-dried and dry affinity chromatography device are summarized in Table M. The dry affinity chromatography devices were wetted using 50 mM sodium phosphate and 150 mM sodium chloride buffer using the methods described above and subsequently tested to assess their liquid permeability and 20 second residence time dynamic binding capacity using the protocols described in the test methods. The performance of these devices is reported in Table N.

[0217] Table M

[0218]

[0219] Table N

[0220]

[0221]

[0222] Device T was further characterized for elution volume performance using a method for determining the elution volume of the device preparation from 100 mAU to 100 mAU. The performance of Device T is shown in Table O. Figure 9 The UV absorbance chromatogram at 280 nm associated with device T characterization using the method for determining the elution volume from 100 mAU to 100 mAU for the device preparation is shown in FIG.

[0223] Table O

[0224]

[0225] The invention of this application has been described above generally and with reference to specific embodiments. It will be apparent to those skilled in the art that various modifications and variations may be made to the embodiments described herein without departing from the scope of the invention. Therefore, the embodiments are intended to cover such modifications and variations of the invention as come within the scope of the appended claims and their equivalents.

Claims

1. A composite membrane for an affinity chromatography device, comprising: a dry fibrillated polymer film having inorganic particles therein, at least one of the inorganic particles and the dry fibrillated polymer film having an affinity ligand covalently bonded thereto, the affinity ligand reversibly binding to a target molecule, wherein the dry fibrillated polymer film has a moisture content of less than or equal to 60% by mass of the dry fibrillated film. 2 . The composite film of claim 1 , wherein the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

3. The composite membrane of claim 1 or claim 2, wherein the inorganic particles have a spherical shape and a nominal particle size of about 5 microns to about 20 microns. 4 . The composite film according to claim 1 , wherein the inorganic particles have a particle size distribution D90 / D10 of 3 or less.

5. The composite membrane of any one of claims 1 to 4, wherein the dry fibrillated polymer membrane comprises an expanded polytetrafluoroethylene membrane, an expanded modified polytetrafluoroethylene membrane, an expanded tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane.

6. The composite membrane of any one of claims 1 to 5, wherein the affinity ligand is selected from the group consisting of protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, oligonucleotides, and combinations thereof. 7 . The composite membrane according to claim 1 , wherein the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

8. The composite film of any one of claims 1 to 7, wherein the dry fibrillated polymer film has a wound configuration.

9. The composite membrane of any one of claims 1 to 8, wherein the dry fibrillated polymer membrane has a stacked configuration.

10. The composite membrane of any one of claims 1 to 9, wherein the dry fibrillated polymer membrane is treated with ethylene oxide, gamma irradiation, or X-ray irradiation for sterilization.

11. Affinity chromatography device, comprising: fluid inlet; a fluid outlet fluidly connected to the fluid inlet; a dry fibrillated polymeric membrane positioned between the fluid inlet and the fluid outlet and containing at least one inorganic particle therein; in, At least one of the following: Before drying, the moisture content of the dry fibrillated polymer film is less than or equal to 60% by mass of the dry fibrillated polymer film, or The water content of the affinity chromatography device is less than or equal to 60% by mass of the affinity chromatography device; and At least one of the dry fibrillated polymer film and the inorganic particles has an affinity ligand covalently bonded thereto, and the affinity ligand reversibly binds to a target molecule.

12. The device of claim 11, comprising a housing surrounding the fluid inlet, the fluid outlet, and the dry fibrillated polymer membrane.

13. A device as claimed in claim 11 or claim 12, wherein two or more of the dry fibrillated membranes are in the form of a dry stacked membrane assembly positioned between the fluid inlet and fluid outlet.

14. A device as claimed in claim 11 or claim 12, wherein the dry fibrillated film has a wound film construction comprising a dry fibrillated polymer wrapped around a core.

15. The device according to any one of claims 11 to 14, wherein the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

16. The device of any one of claims 11 to 15, wherein the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

17. The device of any one of claims 11 to 16, wherein the dry fibrillated polymer film comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

18. The device of any one of claims 11 to 17, wherein the affinity ligand is selected from the group consisting of protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, and combinations thereof.

19. The device of any one of claims 11 to 18, wherein at least one of the dry fibrillated polymer membrane and the affinity chromatography device is treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

20. The device of any one of claims 11 to 19, wherein the affinity chromatography device is configured to achieve a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds.

21. The device of any one of claims 11 to 20, wherein the affinity chromatography device is configured to achieve about 100 (X 10 -12 cm 2 ) to about 2000(X 10 -12 cm 2 ) hydraulic permeability.

22. The device of any one of claims 11 to 21, wherein the affinity chromatography device is configured to achieve an elution volume of from 100 mAU to 100 mAU, 1 column volume (CV) to 6 column volumes (CV).

23. The device of any one of claims 11 to 22, wherein the affinity chromatography device is configured to achieve a cycling durability of at least 100 cycles at an operating pressure of less than 0.3 MPa.

24. Use of the article of any one of claims 11 to 23 for separating target molecules from a fluid stream.

25. A manifold comprising at least two affinity chromatography devices according to any one of claims 11 to 23 arranged in a parallel configuration.

26. The header of claim 25, wherein the header is enclosed in a housing.

27. A device comprising a first manifold and a second manifold in a parallel configuration, wherein each of the first manifold and the second manifold comprises at least two affinity chromatography devices according to any one of claims 11 to 23.

28. The device of claim 27, wherein the first header and the second header are enclosed in a housing.

29. A method for isolating a target molecule, the method comprising: Provided is a chromatographic device comprising: Entrance; export; and a dry fibrillated polymer film having a moisture content of less than or equal to 60% by mass of the dry fibrillated polymer film, the dry fibrillated polymer film being positioned between the inlet and the outlet and containing at least one inorganic particle therein; wetting the dry fibrillated polymer film to form a wet polymer film; adding an aqueous mixture containing a target molecule therein to the inlet; passing the aqueous mixture through a wet fibrillated polymer membrane, wherein the target molecule is bound to the affinity ligand, and allowing the target molecule to bind to an affinity ligand, wherein at least one of the dry fibrillated polymer film and the inorganic particles has the affinity ligand thereon.

30. The method of claim 29, comprising treating the dry fibrillated polymer film with ethylene oxide, thereby sterilizing the dry fibrillated polymer film.

31. The method of claim 30, wherein the treating is performed according to the ethylene oxide (EO) treatment method for films described in the Test Methods section.

32. The method of claim 29, comprising treating the chromatographic device with ethylene oxide, gamma irradiation, or X-ray irradiation.

33. The method of claim 29, comprising sterilizing the chromatography device by treating the chromatography device with ethylene oxide, gamma irradiation, or X-ray irradiation.

34. The method of claim 29, comprising treating a non-dried fibrillated membrane to covalently bond an affinity ligand to one or both of the non-dried fibrillated polymer membrane and inorganic particles.

35. The method of claim 29, comprising characterizing the moisture content of the dry fibrillated polymer film.

36. The method of claim 35, wherein the characterization is performed according to the method for determining the moisture content of the membrane described in the Test Methods section.

37. The method of claim 29, comprising characterizing the moisture content of the chromatographic device.

38. The method of claim 29, comprising drying a non-dried fibrillated film by freeze drying to form the dried fibrillated polymer film.

39. The method of claim 38, wherein the drying is performed according to the membrane freeze-drying method described in the Test Methods section.

40. The method of claim 29, comprising determining the dynamic binding capacity at 10% breakthrough.

41. The method of claim 40, wherein said determining is performed according to the method for determining the dynamic binding capacity of a device article at 10% breakthrough as described in the Test Methods section.

42. The method of claim 29, comprising initially wetting the dry chromatography device.

43. The method of claim 42, wherein said initial wetting is performed according to the method for initial wetting of a dry device article as described in the Test Methods section.

44. The method of claim 29, comprising determining an elution volume of the chromatography device.

45. The method of claim 44, wherein the determining is performed according to the method for determining the elution volume of the device preparation from 100 mAU to 100 mAU described in the Test Methods section.

46. ​​The method of claim 29, comprising determining the dynamic binding capacity of the chromatography device at 10% breakthrough.

47. The method of claim 46, wherein said determining is performed according to the method for determining the dynamic binding capacity of a device article at 10% breakthrough described in the Test Methods section.

48. The method of claim 29, wherein the chromatography device is an affinity chromatography device.

49. The method of claim 29, wherein the affinity ligand is selected from the group consisting of protein A, protein G, protein L, human Fc receptor, protein antibody, polysaccharide, oligosaccharide, and combinations thereof.

50. The method of claim 29, wherein the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

51. The method of claim 29, wherein the dry fibrillated polymer comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

52. Affinity chromatography device, comprising: shell member; allowing fluid to flow into the inlet of the housing member; an outlet for allowing fluid to flow out of the housing member and being fluidly connected to the inlet; a stacked membrane assembly positioned within the housing member between the fluid inlet and the fluid outlet, the stacked membrane assembly comprising: Two or more dry fibrillated polymer films in a stacked configuration, each of the dry fibrillated polymer films containing inorganic particles; wherein the moisture content of the dry fibrillated polymer film is less than or equal to 60% by mass of the dry fibrillated polymer film; and At least one of the dry fibrillated polymer film and the inorganic particles has an affinity ligand covalently bonded thereto, and the affinity ligand reversibly binds to a target molecule.

53. The device of claim 52, wherein the target molecule comprises: a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

54. The device of claim 51 or claim 52, wherein the inorganic particles are selected from the group consisting of spherical particles, non-spherical particles, and combinations thereof.

55. The device of any one of claims 52 to 54, wherein the inorganic particles have a spherical shape and a nominal particle size of about 5 microns to about 20 microns.

56. The device of any one of claims 52 to 55, wherein the particle size distribution has a D90 / D10 of less than or equal to 3.

57. The device of any one of claims 52 to 56, wherein the dry fibrillated polymer film comprises an expanded polytetrafluoroethylene film, an expanded modified polytetrafluoroethylene film, an expanded tetrafluoroethylene copolymer film, or an expanded polyethylene film.

58. The device of any one of claims 52 to 57, wherein the affinity ligand is selected from the group consisting of protein A, protein G, protein L, human Fc receptor protein, antibodies, polysaccharides, oligosaccharides, oligonucleotides, and combinations thereof.

59. The device of any one of claims 52 to 58, wherein the device is treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

60. The device of any one of claims 52 to 59, wherein the dry fibrillated polymer film has a stacked configuration having two sides and a plurality of edges of the stacked film assembly, and wherein the device further comprises a material that seals the edges of the stacked film assembly.

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