Dry fibrillated membrane and affinity chromatography device containing the dry fibrillated membrane

Abstract

The present disclosure relates to affinity chromatography devices including dry fibrillated polymer membranes having a moisture content of 60% or less by weight of the dry fibrillated membrane. Drying of non-dry fibrillated polymer membranes by lyophilization is possible during formation of the dry fibrillated polymer membrane. Dry affinity chromatography devices can improve the stability or viability of purified target molecules, reduce transportation risks (e.g., reduced risk of accidental breakage or leakage due to dropping), and reduce storage risks (e.g., reduced risk of microbial growth during storage). The affinity chromatography device and / or dry fibrillated polymer membrane can be treated with ethylene oxide, gamma irradiation, or X-ray irradiation. In some embodiments, the treatment sterilizes the chromatography device or dry fibrillated polymer membrane. The dry affinity chromatography device can 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 also to affinity chromatography devices including dry fibrillated polymeric membranes that enable the separation of target molecules from aqueous mixtures. [Background technology]

[0002] Chromatography is commonly used to separate and / or purify molecules of interest, such as proteins, nucleic acids, and polysaccharides, from a mixture. In affinity chromatography, the mixture is passed through a matrix to which a ligand (i.e., a specific binding partner) specific for the molecule of interest is attached. Upon contact with the ligand, the molecule of interest binds to the matrix and is therefore removed from the mixture. Affinity chromatography offers several advantages over other types of chromatography. For example, affinity chromatography provides a purification method that can isolate a target protein from a mixture of the target protein and other biomolecules in a single step with high yield. Summary of the Invention [Problem to be solved by the invention]

[0003] Despite the recognized 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, that provide equivalent or better binding capacities, that are reusable, ready to use, and / or ready to sterilize, and that have reduced risks associated with transportation and / or storage. [Means for solving the problem]

[0004] According to one embodiment ("Embodiment 1"), a composite membrane for an affinity chromatography device includes a dry fibrillated polymer membrane having inorganic particles therein, at least one of the inorganic particles and the dry fibrillated polymer membrane having covalently attached thereto an affinity ligand that reversibly binds to a target molecule, and the dry fibrillated polymer membrane has a water content of 60% or less by weight of the dry fibrillated membrane.

[0005] According to another embodiment (embodiment 2), in addition to embodiment 1, the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

[0006] According to another embodiment (embodiment 3), in addition to embodiment 1 or embodiment 2, the inorganic particles have a spherical shape and a nominal particle size of about 5 micrometers to about 20 micrometers.

[0007] According to another embodiment (embodiment 4), in addition to any one of embodiments 1 to 3, the inorganic particles have a particle size distribution D90 / D10 of 3 or less.

[0008] According to another embodiment (Embodiment 5), in addition to any one of Embodiments 1 to 4, 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.

[0009] According to another embodiment (embodiment 6), in addition to any one of embodiments 1 to 5, the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, an oligonucleotide, and combinations thereof.

[0010] According to another embodiment (embodiment 7), in addition to any one of embodiments 1 to 6, the target molecule comprises a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0011] According to another embodiment (embodiment 8), in addition to any one of embodiments 1 to 7, the dry fibrillated polymer membrane has a rolled configuration.

[0012] According to another embodiment (embodiment 9), in addition to any one of embodiments 1 to 8, the dry fibrillated polymer membrane has a stacked configuration.

[0013] According to another embodiment (embodiment 10), in addition to any one of embodiments 1 to 9, the dry fibrillated polymer membrane is sterilized by ethylene oxide, gamma irradiation, or X-ray irradiation.

[0014] According to one embodiment ("Embodiment 11"), an affinity chromatography device includes a fluid inlet, a fluid outlet fluidly connected to the fluid inlet, and a dry fibrillated polymer membrane disposed between the fluid inlet and the fluid outlet and including at least one inorganic particle therein, wherein at least one of (1) the dry fibrillated polymer membrane has a water content of 60% or less by weight of the dry fibrillated polymer membrane before drying, or (2) the affinity chromatography device has a water content of 60% or less by weight of the affinity chromatography device, and at least one of the dry fibrillated polymer membrane and the inorganic particle is covalently bound to an affinity ligand that reversibly binds to a target molecule.

[0015] According to another embodiment (embodiment 12), in addition to embodiment 11, there is included a housing enclosing the fluid inlet, the fluid outlet, and the dry-fibrillated polymer membrane.

[0016] According to another embodiment (embodiment 13), in addition to embodiment 11 or embodiment 12, the two or more dry fibrillated membranes are in the form of a dry stack membrane assembly disposed between the fluid inlet and the fluid outlet.

[0017] According to another example (example 14), in addition to example 11 or example 12, the dry fibrillated membrane has a wound membrane configuration including a dry fibrillated polymer wound around a core.

[0018] According to another embodiment (embodiment 15), in addition to any one of embodiments 11 to 14, the target molecule comprises a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0019] According to another embodiment (embodiment 16), in addition to any one of embodiments 11 to 15, the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

[0020] According to another embodiment (embodiment 17), in addition to any one of embodiments 11 to 16, the dry-laid fibrillated polymer membrane comprises an expanded polytetrafluoroethylene membrane, an expanded modified polytetrafluoroethylene membrane, an expanded tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane.

[0021] According to another embodiment (embodiment 18), in addition to any one of embodiments 11 to 17, the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, and combinations thereof.

[0022] According to another embodiment (embodiment 19), in addition to any one of embodiments 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 another embodiment (embodiment 20), in addition to any one of embodiments 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 another embodiment (embodiment 21), in addition to any one of embodiments 11 to 20, the affinity chromatography device comprises about 100 (×10 -12 cm 2 ) ~ Approximately 2000 (×10 -12 cm 2 ) is configured to achieve a water permeability of 100%.

[0025] According to another embodiment (embodiment 22), in addition to any one of embodiments 11 to 21, the affinity chromatography device is configured to achieve an elution volume of 100 mAU to 100 mAU in 1 column volume (CV) to 6 column volumes (CV).

[0026] According to another embodiment (embodiment 23), in addition to any one of embodiments 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 another embodiment (embodiment 24), in addition to any one of embodiments 11 to 23, the article is used for separating target molecules from a fluid stream.

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

[0029] According to another embodiment (embodiment 26), in addition to embodiment 25, the manifolds are housed in one housing.

[0030] According to another embodiment (embodiment 27), an apparatus comprises a first manifold and a second manifold arranged in a parallel configuration, each of the first manifold and the second manifold comprising at least two affinity chromatography devices according to any one of embodiments 11 to 23.

[0031] According to another embodiment (embodiment 28), in addition to embodiment 27, the first manifold and the second manifold are housed in one housing.

[0032] According to another embodiment (Embodiment 29), a method for isolating a target molecule includes the steps of: (1) providing a chromatography device including an inlet, an outlet, and a dry fibrillated polymer membrane having a water content of 60% or less by weight of the dry fibrillated polymer membrane, wherein the dry fibrillated polymer membrane is disposed between the inlet and the outlet and includes at least one inorganic particle therein; (2) wetting the dry fibrillated polymer membrane to form a wet polymer membrane; (3) adding an aqueous mixture including the target molecule therein to the inlet; (4) passing the aqueous mixture through the wet fibrillated polymer membrane, wherein the target molecule binds to an affinity ligand; and (5) binding the target molecule to the affinity ligand, wherein the affinity ligand is present on at least one of the dry fibrillated polymer membrane and the inorganic particle.

[0033] According to another embodiment (embodiment 30), in addition to embodiment 29, the method includes sterilizing the dry fibrillated polymer membrane by treating the dry fibrillated polymer membrane with ethylene oxide.

[0034] According to another embodiment (Embodiment 31), in addition to embodiment 30, the treatment is carried out according to the membrane ethylene oxide (EO) treatment method described in the Test Methods section.

[0035] According to another embodiment (embodiment 32), in addition to embodiment 29, the method includes treating the chromatographic device with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0036] According to another embodiment (embodiment 33), in addition to embodiment 29, the method includes sterilizing the chromatography device by treating the chromatography device with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0037] According to another embodiment (embodiment 34), in addition to embodiment 29, the method includes treating the non-dry fibrillating membrane to covalently bond affinity ligands to one or both of the non-dry fibrillating polymer membrane and the inorganic particles.

[0038] According to another embodiment (embodiment 35), in addition to embodiment 29, the method includes the step of characterizing the moisture content of the dry fibrillated polymer membrane.

[0039] According to another embodiment (Embodiment 36), in addition to embodiment 35, the characterization is performed according to the method for determining membrane moisture content described in the Test Methods section.

[0040] According to another embodiment (embodiment 37), in addition to embodiment 29, further comprising the step of characterizing the water content of the chromatography device.

[0041] According to another embodiment (embodiment 38), in addition to embodiment 29, the method includes the step of drying the non-dry fibrillated membrane by lyophilization to form a dry fibrillated polymer membrane.

[0042] According to another embodiment (embodiment 39), in addition to embodiment 38, drying is carried out according to the membrane freeze-drying method described in the Test Methods section.

[0043] According to another embodiment (embodiment 40), in addition to embodiment 29, the method includes determining the dynamic binding capacity at 10% breakthrough.

[0044] According to another embodiment (Embodiment 41), in addition to embodiment 40, the determination is carried out according to the method for determining the dynamic binding capacity at 10% breakthrough of an article of equipment described in the Test Methods section.

[0045] According to another embodiment (embodiment 42), in addition to embodiment 29, the method includes the step of first wetting the dry chromatography apparatus.

[0046] According to another embodiment (Embodiment 43), in addition to embodiment 42, the initial wetting step is performed according to the Dry Equipment Article Incipient Wetness Method described in the Test Methods section.

[0047] According to another embodiment (embodiment 44), in addition to embodiment 29, the method comprises determining an elution volume of the chromatography device.

[0048] According to another embodiment (Embodiment 45), in addition to embodiment 44, the determination is carried out according to the method for determining the elution volume of an article of equipment from 100 mAU to 100 mAU described in the Test Methods section.

[0049] According to another embodiment (embodiment 46), in addition to embodiment 29, the method includes determining the dynamic binding capacity at 10% breakthrough of the chromatographic device.

[0050] According to another embodiment (Embodiment 47), in addition to embodiment 46, the determination is carried out according to the method for determining the dynamic binding capacity at 10% breakthrough of an article of equipment described in the Test Methods section.

[0051] According to another embodiment (embodiment 48), in addition to embodiment 29, the chromatography device is an affinity chromatography device.

[0052] According to another embodiment (embodiment 49), in addition to embodiment 29, the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, and combinations thereof.

[0053] According to another embodiment (embodiment 50), in addition to embodiment 29, the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

[0054] According to another embodiment (embodiment 51), in addition to embodiment 29, the dry fibrillating polymer comprises an expanded polytetrafluoroethylene membrane, an expanded modified polytetrafluoroethylene membrane, an expanded tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane.

[0055] According to another aspect (Aspect 52), an affinity chromatography device includes a housing member, an inlet that allows fluid to enter the housing member, an outlet that allows fluid to exit the housing member and is fluidly connected to the inlet, and a stack membrane assembly disposed within the housing member between the fluid inlet and the fluid outlet, wherein the stack membrane includes two or more dry fibrillated polymer membranes in a stack configuration, each of the dry fibrillated polymer membranes containing inorganic particles therein, the dry fibrillated polymer membranes having a water content of 60% by weight or less of the dry fibrillated polymer membrane, and at least one of the dry fibrillated polymer membrane and the inorganic particles is covalently bound to an affinity ligand that reversibly binds to a target molecule.

[0056] According to another embodiment (embodiment 53), in addition to embodiment 52, the target molecule comprises a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

[0057] According to another embodiment (embodiment 54), further to embodiment 51 or embodiment 52, the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

[0058] According to another embodiment (embodiment 55), in addition to any one of embodiments 52 to 54, the inorganic particles have a spherical shape and a nominal particle size of about 5 micrometers to about 20 micrometers.

[0059] According to another embodiment (embodiment 56), in addition to any one of embodiments 52 to 55, the particle size distribution has a D90 / D10 of 3 or less.

[0060] According to another embodiment (embodiment 57), in addition to any one of embodiments 52 to 56, the dry-laid fibrillated polymer membrane comprises an expanded polytetrafluoroethylene membrane, an expanded modified polytetrafluoroethylene membrane, an expanded tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane.

[0061] According to another embodiment (embodiment 58), in addition to any one of embodiments 52 to 57, the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, an oligonucleotide, and combinations thereof.

[0062] According to another embodiment (embodiment 59), in addition to any one of embodiments 52 to 58, the device has been treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

[0063] According to another embodiment (embodiment 60), in addition to any one of embodiments 52 to 59, the dry fibrillated polymer membrane has a stack configuration having two sides of a stack membrane assembly and a plurality of edges, and the device further includes a material that seals the edges of the stack membrane assembly.

[0064] The foregoing examples are illustrative only and should not be construed as limiting or narrowing the scope of the inventive concepts otherwise provided by this disclosure. While multiple examples are disclosed, still other embodiments will become apparent to those skilled in the art from the following detailed description. The following detailed description shows and describes examples for illustrative purposes. Accordingly, the drawings and detailed description are to be regarded as illustrative in nature and not restrictive. [Brief explanation of the drawings]

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

[0066] [Figure 1] FIG. 1 is an exploded view of a chromatography device including a wound membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein, according to at least one embodiment.

[0067] [Figure 2]FIG. 1 is a cross-sectional view of a chromatography device including a wound membrane assembly showing a dry fibrillated polymer membrane, an outer flow channel, and an inner flow channel, according to at least one embodiment.

[0068] [Figure 3] FIG. 1 is an exploded view of a chromatography device including a stack membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein, according to at least one embodiment.

[0069] [Figure 4A] FIG. 1 is a cross-sectional view of a chromatography device including a stack membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein, according to at least one embodiment.

[0070] [Figure 4B] FIG. 4B is an exploded view of the chromatography device of FIG. 4A according to at least one embodiment.

[0071] [Figure 5] FIG. 1 is a side view of a manifold containing multiple chromatography devices in a parallel configuration, where each chromatography device includes a stack membrane assembly including a dry-fibrillated polymer membrane having inorganic particles therein, according to at least one embodiment.

[0072] [Figure 6] FIG. 1 is a schematic diagram of a side view of two manifolds in a side-by-side configuration according to at least one embodiment.

[0073] [Figure 7] FIG. 1 is a side view of a manifold including multiple chromatography devices in a parallel configuration, where each chromatography device includes a wound membrane assembly including a dry fibrillated polymer membrane having inorganic particles therein, according to at least one embodiment.

[0074] [Figure 8]FIG. 1 is a side view of two manifolds in a side-by-side configuration including a wound dry fibrillated polymer membrane assembly according to at least one embodiment.

[0075] [Figure 9] 1 is a chromatogram of UV absorbance at 280 nm associated with characterization of Device T, according to at least one embodiment.

[0076] [Figure 10A] FIG. 1 is a schematic diagram of a multi-well plate according to at least one embodiment.

[0077] [Figure 10B] FIG. 10B is a schematic diagram of a portion of the multiwell plate shown in FIG. 10A, showing a portion of a stacked membrane assembly disposed on a porous substrate, according to at least one embodiment. DETAILED DESCRIPTION OF THE INVENTION

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

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

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

[0081] As used herein, the term "fibrillated polymer membrane" is meant to refer to the presence of fibrils within a polymer membrane, for example, a polymer membrane having a microstructure characterized by nodes, fibrils, and voids, where the nodes are interconnected by fibrils and the voids are the spaces located between the nodes and fibrils.

[0082] The terms "fibrillated polymer membrane," "polymer membrane," and "membrane" may be used interchangeably herein.

[0083] As used herein, the term "spiral wound membrane assembly" is meant to include both a single dry fibrillated polymer membrane and a dry fibrillated polymer membrane containing one or more intermediate materials in a spiral configuration.

[0084] As used herein, the term "stack membrane assembly" is meant to include both standalone dry fibrillated polymer membranes and dry fibrillated polymer membranes containing 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" can be used interchangeably.

[0086] The term "wet fibrillated polymeric membrane" as used herein is meant to refer to a dry fibrillated membrane or dry fibrillated membrane assembly that has been subjected to a wetting process.

[0087] The terms "non-dry," "wet," and "moist" are used interchangeably herein and are meant to refer to something prior to undergoing a drying process to become a dry fibrillated membrane or dry chromatography device.

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

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

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

[0091] The present disclosure is directed to a dry fibrillated polymer membrane having a water content of 60% or less by weight of the dry membrane. The dry fibrillated polymer membrane can be arranged in a wound or stacked configuration within an affinity chromatography device. The affinity chromatography device (including the dry fibrillated polymer membrane) can have a water content of 60% or less by weight of the device. Additionally, the dry fibrillated polymer membrane includes inorganic particles (e.g., spherical or irregularly shaped) within and / or on the membrane. Affinity ligands can be bound to the inorganic particles and / or the dry fibrillated polymer membrane. When forming the dry fibrillated polymer membrane, the non-dry fibrillated polymer membrane can be dried by lyophilization. This drying process by lyophilization allows for a more simplified device manufacturing process, in which chemical immobilization of ligands can be performed on the fibrillated membrane prior to integration into the chromatography device, thereby eliminating an in-situ chemical immobilization step from the device manufacturing process.

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

[0093] The chromatography device (in any form, including but not limited to, an affinity chromatography device, a cassette, a cassette assembly, a manifold, a manifold chromatography device, a branched manifold, and / or a branched manifold chromatography device, and any other chromatography device described herein) comprises no more than 60% by weight of the dry affinity chromatography device, or less than about 55% by weight of the dry device, or less than about 50% by weight of the dry device, or less than about 45% by weight of the dry device. %, or less than about 40% by weight of the dry equipment, or less than about 35% by weight of the dry equipment, or less than about 30% by weight of the dry equipment, or less than about 25% by weight of the dry equipment, or less than about 20% by weight of the dry equipment, or less than about 15% by weight of the dry equipment, or less than about 10% by weight of the dry equipment, or less than about 5% by weight of the dry equipment, or less than about 3% by weight of the dry equipment, or less than about 1% by weight of the dry equipment, or less than about 0.1% by weight of the dry equipment, or less than about 0.01% by weight of the dry equipment, or less than about 0.001% by weight of the dry equipment, or 0% by weight of the equipment.

[0094] The dry fibrillated polymer membrane (in any form, including but not limited to, a fibrillated polymer membrane, a fibrillated membrane disk, a stacked membrane assembly, a wound membrane assembly, a spiral wound membrane assembly, a membrane assembly, and / or a fibrillated membrane layer, and any other fibrillated polymer membrane described herein) comprises no more than 60% by weight of the dry fibrillated polymer membrane, or less than about 55% by weight of the dry membrane, or less than about 50% by weight of the dry membrane, or less than about 45% by weight of the dry membrane, or may have a moisture content of less than about 40% by weight of the dry film, or less than about 35% by weight of the dry film, or less than about 30% by weight of the dry film, or less than about 25% by weight of the dry film, or less than about 20% by weight of the dry film, or less than about 15% by weight of the dry film, or less than about 10% by weight of the dry film, or less than about 5% by weight of the dry film, or less than about 3% by weight of the dry film, or less than about 1% by weight of the dry film, or less than about 0.1% by weight of the dry film, or less than about 0.01% by weight of the dry film, or less than about 0.001% by weight of the dry film, or 0% by weight of the dry film.

[0095] The dry fibrillated polymer membrane (in any form, including but not limited to, a fibrillated polymer membrane, a fibrillated membrane disk, a stacked membrane assembly, a wound membrane assembly, a spiral wound membrane assembly, a membrane assembly, and / or a fibrillated membrane layer, and any other fibrillated polymer membrane described herein) comprises between about 0.001% and 60% by weight of the dry fibrillated polymer membrane, or between about 0.001% and about 55% by weight of the dry membrane. %, or about 0.01% to about 50% 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 membrane has a moisture content of about 0.001% to about 5% by weight of the dry fibrillated polymer membrane, or about 0.01% to about 4.5% by weight of the dry membrane, or about 0.1% to about 4% by weight of the dry membrane, or about 1.0% to about 3.5% by weight of the dry membrane, or about 1.1% to about 3% by weight of the dry membrane. In some embodiments, the dry fibrillated polymer membrane has a moisture content of about 1.3% to about 2% by weight of the dry membrane.

[0096] The dry chromatography device (in any form, including but not limited to, an affinity chromatography device, a cassette, a cassette assembly, a manifold, a manifold chromatography device, a branched manifold, and / or a branched manifold chromatography device, and any other chromatography device described herein) comprises between about 0.001% and 60% by weight of the dry affinity chromatography device, or about 0.001% by weight of the dry device. % to about 55% by weight of the dry equipment, or about 0.01% to about 50% by weight of the dry equipment, or about 0.05% to about 45% by weight of the dry equipment, or about 0.1% to about 40% by weight of the dry equipment, or about 0.5% to about 35% by weight of the dry equipment, or about 0.6% to about 30% by weight of the dry equipment, or about 0.7% to about 25% by weight of the dry equipment, or about 0.8% to about 20% by weight of the dry equipment, or about 0.9% to about 15% by weight of the dry equipment, or about 1% to about 10% by weight of the dry equipment. In some embodiments, the chromatography device has a moisture content of about 0.001% to about 5% by weight of dry chromatography device, or about 0.01% to about 4.5% by weight of dry device, or about 0.1% to about 4% by weight of dry device, or about 1.0% to about 3.5% by weight of dry device, or about 1.1% to about 3% by weight of dry device. In some embodiments, the chromatography device has a moisture content of about 1.3% to about 2% by weight of dry device.

[0097] In some embodiments, the chromatography device has a dynamic binding capacity (DBC) of greater than 35 mg / ml at 10% breakthrough with a 20 second residence time. Furthermore, the affinity chromatography device can have a cycling durability of at least 100 cycles without exceeding an operating pressure of 0.3 MPa. In further embodiments, the affinity chromatography device has an elution volume of 100 mAU to 100 mAU, which is about 1 column volume to about 6 column volumes. In some embodiments, dry affinity chromatography devices can improve the stability or viability of the target molecule being purified. According to some embodiments, dry affinity chromatography devices can reduce storage risks because the elimination of fluids within the device reduces the risk of microbial growth during storage. According to some embodiments, dry affinity chromatography devices can reduce transportation risks because the elimination of fluids within the device reduces the risk of accidental breakage or leakage due to dropping. In some embodiments, affinity chromatography devices including dry fibrillated polymer membranes can also reduce transportation and flow-through lead times. After storage and / or transport, the dry affinity chromatography device can then be easily wetted for use with bioprocessing fluids.

[0098] In some embodiments, dry affinity chromatography devices can have higher permeability and / or long-term stability at ambient conditions. Affinity chromatography devices according to the present disclosure can include other advantages, such as uniform flow fronts under normal flow in multiple configurations (e.g., stacks, spiral wound, cassettes, and parallel-branched configurations), allowing users to incorporate multiple dry affinity chromatography devices in parallel-branched configurations, and / or scalability of performance through residence time. The terms "manifold" and "parallel-branched" are used interchangeably herein and should be understood to define the same configuration. Dry affinity chromatography devices incorporating dry fibrillated polymer membranes allow for various processing modes of the final device (e.g., sterilizability by ethylene oxide treatment, gamma treatment, X-ray treatment, etc., before or after assembly). Such sterilization methods are more limited or impossible when working with existing wet-article device configurations.

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

[0100] 1 and 2 are exploded and cross-sectional views, respectively, of a chromatography device 100 including a wound membrane according to at least one embodiment. In forming the chromatography device 100, in some embodiments, at least one inner intermediate material 200 can be circumferentially disposed on (e.g., wrapped around) a cylindrical core 150 to reach a desired width or pre-specified volume. A dry fibrillated polymer membrane 210 containing spherical and / or non-spherical inorganic particles therein is then wound around the core 150 over the inner intermediate material 200 to reach the desired width or pre-specified volume. An outer layer of at least one outer intermediate material 220 is then circumferentially disposed on (e.g., wrapped around) the dry fibrillated polymer membrane 210 to reach the desired width or pre-specified volume. The combination of the inner intermediate material 200, dry fibrillated polymer membrane 210, and outer intermediate material 220 is referred to herein as a "wound membrane assembly." In some embodiments, a "wound membrane assembly" can also include any combination of polymer and / or polymer intermediate material wound around a cylindrical core. The cylindrical core 150 can have a hollow or solid interior. In either case, the core 150 includes a solid outer wall such that the aqueous mixture flowing through the chromatography device 100 flows through the inner flow channel 140 formed by the inner intermediate material 200 and not within the core 150. The use of a hollow core 150 reduces the amount of material used to form the core 150, reducing the weight of the device 100 and reducing manufacturing costs.

[0101] As shown in FIG. 1 , the wound membrane assembly 110 includes an inner intermediate material 200, a dry-fibrillated polymer membrane 210, and an outer intermediate material 220. The wound membrane assembly 110 and central core 150 can be disposed within a housing 50. In some embodiments, the housing 50 is cylindrical. As shown in FIG. 2 , the outer intermediate material 220 forms the outer flow channel 130, and the inner intermediate material 200 forms the inner flow channel 140. It should be understood that the intermediate materials 200, 220 in the embodiments described herein can be different or the same. Furthermore, two or more intermediate materials can be used to form one or both of the outer flow channel 130 and the inner flow channel 140. Non-limiting examples of materials suitable for forming the housing of the chromatography devices 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). Furthermore, the shape of the housing unit is not limited and can be any shape that encapsulates the affinity chromatography device and / or cassettes (described below) and / or manifolds (described below).

[0102] 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 within the inlet cap 60. The mixture flows through the distributor cap 65 and is directed toward the outer flow channels 130 formed by the outer intermediate material 220. The distributor cap 65 directs the aqueous mixture toward the outer flow channels 130 (i.e., the intermediate material 220) at an angle of approximately 90 degrees from the feed direction. This redirection promotes more uniform flow of the aqueous mixture into the outer flow channels 130. The outer intermediate material 220, which forms the outer flow channels 130, is located between the housing 50 and the wound dry fibrillated polymer membrane 210. The distributor cap 65 may be polyolefin or may be coated with a polyolefin.

[0103] The aqueous mixture flows through the outer flow channel 130 (i.e., the outer intermediate material 220) in a normal direction (e.g., from the outer flow channel 130 to the inner flow channel 140) across the "wet" wound fibrillated polymer membrane 210. As the aqueous mixture passes in normal flow from the outer flow channel 130 (i.e., the outer intermediate material 220) across the wet wound fibrillated polymer membrane 210, the affinity ligands reversibly bind to the target molecules, thereby effectively removing the target molecules from the aqueous mixture. The aqueous mixture, from which the target molecules have 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. The aqueous mixture is then redirected by the outlet cap 75 at the bottom of the inner flow channel 140. The aqueous mixture then flows out of the chromatography device 100 through an outlet 85 located within the outlet cap 75.

[0104] By adjusting 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, larger volumes can be achieved without adversely affecting the performance of the chromatographic device. Additionally, as known to those skilled in the art, target molecules can be removed from affinity ligands, for example, by passing a fluid having a lower pH through the chromatographic device.

[0105] The intermediate materials 200 and 220 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 films, or porous non-fluoropolymer films (e.g., porous polypropylene or other porous polyolefin films), porous nonwoven materials, or porous woven materials. In some embodiments, the intermediate film is a thermoplastic or thermosetting polymer film. In some embodiments, the wound membrane assembly includes an integrated inlet end cap 60 and / or an integrated outlet end cap 75 at opposite ends of the housing 50, thereby forming an integrated, reusable chromatography device. In some embodiments, the inlet or inlet end cap 60 and the outlet or outlet end cap 75 can 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 results in sterilization of the device 100 or membrane 210. In some embodiments, the dry fibrillated polymer membrane 210 can be sterilized separately before 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, as shown generally in FIG. 3 , a chromatography device 300 includes dry fibrillated polymer membranes configured as individual dry fibrillated polymer membrane disks 340 stacked together to form a stacked membrane assembly 320. The dry fibrillated polymer membrane disks 340 can be arranged in a stacked configuration by simply stacking the dry fibrillated polymer membrane disks 340 on top of each other. Alternatively, the dry fibrillated polymer membrane disks 340 can be joined together after stacking using heat and / or pressure or other conventional methods. The stacked membrane assembly 320 described herein refers to dry fibrillated polymer membrane disks for ease of explanation. Dry fibrillated polymer membranes can be formed in other geometric and / or non-geometric shapes, and this is considered within the scope of the present disclosure. It should be understood that when an affinity chromatography device 300 includes a dry fibrillated polymer membrane or dry fibrillated polymer membrane disks 340, the chromatography device 300 is considered “dry.”

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

[0109] The stack membrane assembly 320 can be disposed within 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, when the dry-laid fibrillated polymer membrane has a disk-shaped or circular configuration, the housing 350 is cylindrical; however, any geometric shape capable of housing the stack membrane assembly and achieving the desired dynamic binding capacity is considered within the scope of this 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, so long as the porous intermediate materials 360, 380 do not interfere with the operation of the chromatography device.

[0110] The dry fibrillated polymer membrane disks 340 in the stack membrane assembly 320 can be adhered to the housing 350 at the interior walls of the housing 350 by any conventional process (e.g., melt sealing or use of a sealant) that prevents flow between the periphery of the dry fibrillated polymer membrane disks 340 and the housing 350. The inlet caps 365 and outlet caps 375 can be sealed to the housing 350 by a similar or identical process. The inlet caps 365 and outlet caps 375 include an inlet 380 and an outlet 385, respectively, that allow aqueous mixtures to flow into and out of the affinity chromatography device 300. Specifically, the inlet 380 allows fluid to flow into the housing 350, and the outlet 385 allows fluid to flow out of the housing 350. The dry fibrillated polymer membrane disks 340 remain dry prior to use of the chromatography device 300. Prior to use, the dry affinity chromatography device 300 or dry fibrillated polymer membrane / dry fibrillated membrane disc 340 is wetted with a liquid suitable for bioprocessing, including but not limited to aqueous buffers, harvest, acids, bases, organic solvents, and other fluids commonly used in bioprocessing and known to those skilled in the art.

[0111] In use, an aqueous mixture flows sequentially through the upper intermediate material 360, through the "wet" fibrillated polymer membrane disks 340 that form the stack membrane assembly 320, and through the lower intermediate material 380. As the aqueous mixture passes through the "wet" fibrillated polymer membrane disks 340, the affinity ligands reversibly bind to the target molecules, thereby effectively removing them from the aqueous mixture. As known to those skilled in the art, target molecules can be removed from the affinity ligands by, for example, passing a fluid having a lower pH through the device.

[0112] In some embodiments, the affinity chromatography device 300 and / or the dry fibrillated polymer membrane discs 340 can be treated with ethylene oxide, gamma irradiation, X-ray irradiation, or other treatment methods that can result in sterilization of the device 300 (including sterilization of the dry fibrillated membrane discs 340). In some embodiments, the stack membrane assembly 320 or the dry fibrillated polymer discs can be sterilized with any of the aforementioned sterilization methods prior to assembly into the affinity chromatography device.

[0113] The total number of dry fibrillated polymer membranes present in a stack membrane assembly is not particularly limited and depends on the desired end use and / or desired mass transport flow within the stack membrane assembly. The stack membrane assembly can 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 membranes. It should be understood that there can be hundreds or even thousands of dry fibrillated polymer membranes within a stack membrane assembly.

[0114] As previously mentioned, the dry-laid fibrillated polymer membrane contains inorganic particles therein. In some embodiments, the dry-laid fibrillated polymer membrane can contain one or more types of inorganic particles and / or one or more nominal particle sizes within the dry-laid fibrillated polymer membrane. Non-limiting examples of suitable inorganic particles include, but are not limited to, silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof. As used herein, the term "silica" should be understood to refer to silicon dioxide that does not contain or does not contain measurable amounts of boron, as measured by X-ray photoelectron spectroscopy (XPS). Furthermore, the inorganic particles can be solid or porous and can have a variety of sizes and shapes.

[0115] The inorganic particles can have a nominal particle size of about 0.1 micrometers or greater, about 0.5 micrometers or greater, about 1 micrometer or greater, about 5 micrometers or greater, about 10 micrometers or greater, about 15 micrometers or greater, about 20 micrometers or greater, or about 25 micrometers or greater. Further, the inorganic particles can be monodisperse or polydisperse.

[0116] Both the dry fibrillated polymer membrane of the wound membrane assembly and the dry fibrillated polymer membrane of the stack membrane assembly can contain spherical inorganic particles or particles having a spherical configuration. As used herein, the term "spherical" means that the inorganic particles have a round or nearly round shape, where the distance from the center of the inorganic particle to the outer edge of the particle is the same or nearly the same at any point. In some embodiments, the spherical inorganic particles can have a particle size distribution with a D90 / D10 of 3 or less, 2.5 or less, 2 or less, 1.5 or less, or 1 or less. The spherical inorganic particles have a nominal particle size that can be about 5 micrometers, about 10 micrometers, about 15 micrometers, about 20 micrometers, and combinations and blends thereof. In some embodiments, the spherical inorganic particles are polydisperse.

[0117] In some embodiments, the affinity ligand is covalently bound to the inorganic particles. In another embodiment, the affinity ligand is covalently bound to the dry-fibrillated polymer membrane. In a further embodiment, the affinity ligand can be bound to both the dry-fibrillated polymer membrane and the inorganic particles. The affinity ligand can be a protein, antibody, or polysaccharide that reversibly binds to a target protein or target antibody. In one embodiment, the affinity ligand is, for example, a protein that reversibly binds to 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 polysaccharide that reversibly binds to a specific protein or protein fragment. Affinity ligands used in affinity chromatography devices can include, but are not limited to, protein A, protein G, protein L, human Fc receptor proteins, antibodies that specifically bind to other proteins, and heparin. The affinity ligand can be natural, recombinant, or synthetic. In some embodiments, the affinity ligand may be an oligosaccharide or oligonucleotide that reversibly binds to nucleic acids, including but not limited to mRNA. In some embodiments, the affinity ligand may be an antibody or polysaccharide that reversibly binds to viral vectors, including but not limited to adeno-associated viruses. In yet another embodiment, the affinity ligand is a metal affinity ligand that reversibly binds to His-tagged proteins.

[0118] According to some embodiments, 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. In some embodiments, the dry fibrillated polymer membrane comprises an expanded polytetrafluoroethylene membrane or an expanded polyethylene membrane.

[0119] While embodiments of a wound membrane assembly 110 and a stack membrane assembly 320 are described herein, it should be understood that any number of dry fibrillated polymer membranes and all combinations of dry fibrillated polymer membrane types, spherical inorganic particle types, spherical inorganic particle sizes, and dry fibrillated polymer membrane orientations within the membrane assemblies 110, 320 are within the scope of the present disclosure. Additionally, some or all of the dry fibrillated polymer membranes may differ from one another in terms of composition, thickness, permeability, etc.

[0120] FIG. 4A is a cross-sectional view of a chromatography device 400 including a cassette assembly 401. FIG. 4B is an exploded view of the chromatography device 400 shown in FIG. 4A. The cassette assembly 401 includes a dry fibrillated polymer membrane assembly having inorganic particles therein. The dry fibrillated polymer membrane assembly can be configured to form a stacked membrane assembly 410 by stacking or juxtaposing individual dry fibrillated polymer membrane layers 440. In some embodiments, the dry fibrillated polymer membrane layers 440 can be arranged 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 joined together after stacking (e.g., at intermittent intervals throughout the membrane layers or bonded at the edges of each membrane layer). As shown, affinity chromatography device 400 (e.g., cassette assembly 401) includes a fluid inlet channel 402 connected to an inlet housing 404 and a fluid outlet channel 406 connected to an outlet housing 408. The fluid inlet channel 402 is in fluid communication with the fluid outlet channel 406. In some embodiments, device 400 includes a dry fibrillated polymer membrane assembly 410 including at least one inorganic particle disposed between inlet housing 404 and outlet housing 408. It should be understood that chromatography device 400 is considered "dry" when it includes a dry fibrillated polymer membrane.

[0121] The chromatography device 400 optionally includes at least one inlet intermediate material 460 disposed on the inlet side of the dry stack membrane assembly 410 and at least one outlet intermediate material 480 disposed on the outlet side of the dry stack membrane assembly 410. The inlet intermediate material 460 and the outlet intermediate material 480 may be the same or different. As with the wound membrane and stack membrane assemblies described above, the inlet intermediate material 460 and the outlet intermediate material 480 used to form the cassette stack membrane assembly 410 are not particularly limited, so long as the aqueous mixture can flow through the inlet intermediate material 460 and the outlet intermediate material 480. Non-limiting examples of suitable intermediate materials include, but are not limited to, porous fluoropolymer or non-fluoropolymer films (e.g., porous polypropylene or other porous polyolefin films), porous nonwoven materials, or porous woven materials. In some embodiments (not shown), the inlet intermediate material 460 and the outlet intermediate material 480 are not present.

[0122] In some embodiments, at least one of the dry fibrillated polymer membrane layer 440 and the inorganic particles has covalently attached affinity ligands that reversibly bind to target molecules. 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 that will sterilize the dry fibrillated polymer membrane layer 440 or the chromatography device 400.

[0123] As shown in FIGS. 4A and 4B , the inlet housing 404 and the outlet housing 408 are stacked or arranged side-by-side in a substantially planar manner on opposite sides of the dry stack membrane assembly 410, thereby forming the cassette assembly 401. The dry stack membrane assembly 410 includes a dry fibrillated polymer membrane layer 440. In some embodiments, the inlet housing 404 and the outlet housing 408 have substantially the same shape as the dry fibrillated polymer membrane assembly 410. For example, as shown in FIGS. 4A and 4B , both housings 404, 408 and the dry stack membrane assembly 410 are substantially rectangular in shape. In some embodiments, the housings 404, 408 and the dry stack membrane assembly 410 can be any shape, so long as the housings 404 and 408 completely surround the dry stack membrane assembly 410.

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

[0125] 4B, the inlet intermediate material 460 and the outlet intermediate material 480 can be outside of the peripheral sealing material 412. In some embodiments, the inlet intermediate material 460 and the outlet intermediate material 480 can each comprise a separate intermediate peripheral sealing material that is different from the peripheral sealing material 412 that seals the edges of the stack membrane assembly 410. Additionally, in some embodiments (not shown), the inlet intermediate material 460 and the outlet intermediate material 480 can be inside the peripheral sealing material 412 and thus can be sealed therein.

[0126] 5 is a side view of a parallel branched chromatography device 500 having multiple chromatography devices (each a cassette assembly), each containing a dry fibrillated polymer membrane having inorganic particles therein. In some embodiments, the multiple chromatography devices can contain a mixture of "wet" fibrillated polymer membranes and dry fibrillated polymer membranes.

[0127] As shown, manifold chromatography device 500 includes six cassette assemblies 501a-501f (e.g., cassette assembly 401 in FIG. 4A) arranged in a parallel configuration. In some embodiments, a single manifold can include anywhere from 2 to 20 cassette assemblies arranged parallel to one another. In some embodiments, a single manifold can include more than 20 parallel cassette assemblies, so long as there is a similar distribution of flow and permeability across the cassette assemblies. This similarity across cassette assemblies allows for scalability in device volume and performance.

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

[0129] In some embodiments, manifold chromatography device 500 can include an optional gas separation device 530 (e.g., an integrated air trap) upstream of fluid inlet channel 502. It should be understood that fluid inlet channel 502 and fluid outlet channel 504 can be on opposite sides of the manifold of the chromatography device, on the same side, or in other configurations, so long as fluid flow through the individual chromatography devices 501a-501f occurs in parallel.

[0130] 6 is a cross-sectional view of a branched manifold 650 including at least two manifolds 600A and 600B (e.g., the manifolds of the chromatography device 500 of FIG. 5) disposed within an optional housing 620. Each manifold includes a cassette assembly each including a dry-laid fibrillated polymer membrane having inorganic particles therein.

[0131] As shown in FIG. 6, the branched manifold 650 includes two manifolds 600A and 600B arranged in a parallel configuration (e.g., the manifolds of the chromatography device 500 of FIG. 5). In some embodiments, the branched manifold can include anywhere from 2 to 20 manifolds in parallel with each other. In some embodiments (not shown), the branched manifold can include more than 20 manifolds in parallel with each other. It should be understood that the fluid inlet channel 652 and the fluid outlet channel 654 can be on opposite sides of the manifold of the chromatography device, on the same side, or in other configurations, so long as fluid flow through the individual manifolds 600A and 600B occurs in parallel. In some embodiments, the manifolds 600A and 600B can include fluid outlet channels 654A and 654B, respectively, and optional gas separation devices 630A and 630B (e.g., integrated air traps) upstream of the fluid inlet channels 652A and 652B. In a further embodiment (not shown), an optional gas separation device can be placed upstream of the fluid inlet channel 652 of the branched manifold device.

[0132] 7 is a side cross-sectional view of a manifold 750 including multiple chromatography devices, each including a wound membrane assembly. Each wound membrane assembly includes a dry fibrillated polymer membrane having inorganic particles therein. The chromatography devices are arranged in a parallel configuration. When the chromatography device described with respect to FIG. 7 includes a dry fibrillated polymer membrane or a dry wound membrane assembly, the chromatography device is considered a dry chromatography device.

[0133] In some embodiments, the affinity chromatography devices described herein can be used in a single manifold 750 that includes wound membrane affinity chromatography devices 700, 701 (e.g., the chromatography device 100 shown in FIGS. 1 and 2) arranged in a parallel configuration, as shown schematically in FIG. 7. In some embodiments, the manifold 750 can include an optional gas separation device 730 (e.g., an integrated air trap) upstream of the fluid inlet 740. In some embodiments, the manifold 750 is disposed within a housing 720.

[0134] In use, the dry-stack membrane assemblies or dry-fibrillated polymer membranes disposed within each chromatography device are first wetted with a liquid suitable for bioprocessing. Such liquids include, but are not limited to, aqueous buffers, harvest materials, acids, bases, organic solvents, and other fluids commonly used in bioprocessing and known to those skilled in the art. An aqueous mixture then flows into fluid inlet 740, which splits the aqueous mixture into at least two inlet tubes 760, 761. The split aqueous mixture within inlet tubes 760, 761 flows into chromatography devices 700, 701, where target molecules are captured by affinity ligands disposed on the fibrillated polymer membranes and / or inorganic particles. Aqueous solutions, representing the aqueous mixture minus the target molecules captured by the affinity ligands, exit chromatography devices 700, 701 through outlet tubes 780, 781, respectively. The aqueous solutions within outlet tubes 780, 781 are combined at distribution element 790 and reconstituted into a single aqueous solution.

[0135] The two chromatography devices 700, 701 are shown in FIG. 7 for illustrative purposes only, and it should be understood that two or more of the chromatography devices described herein can be used in a parallel configuration within a single manifold, so long as there is a similar distribution of flow and permeability throughout the chromatography device. This similarity across affinity chromatography devices allows for scalability in device volume and performance. Furthermore, affinity chromatography devices can be incorporated into a parallel configuration system without any modifications or additions to the manifold 750. It should be understood that the fluid inlet 740 and fluid outlet 790 can be on opposite sides of the manifold of the chromatography device, on the same side, or in other configurations, so long as fluid flow through the individual chromatography devices 700, 701 occurs in parallel.

[0136] Figure 8 is a cross-sectional view of two manifolds in a parallel configuration including wound dry-fibrillated polymer membrane assemblies according to at least one embodiment. As shown in Figure 8, the branched manifold 26 includes two manifolds 6a, 6b in a parallel configuration. Manifolds 6a and 6b (e.g., manifold 750 in Figure 7) each include at least two affinity chromatography devices 26a, 26b, and 26c, 26d. The availability of at least two manifolds in parallel advantageously allows for increased volumetric capacity while using the chromatography devices described herein. In some embodiments, the branched manifold 26 can include an optional gas separation device 830 (e.g., an integrated air trap) upstream of the fluid inlet 30.

[0137] 10A and 10B, the dry fibrillated membranes in the form of stacked membrane assemblies described above can be fixed to a multiwell plate 1000 that includes a porous surface 1030 that separates a lower chamber operable at low pressure from an upper chamber operated at high pressure (e.g., atmospheric pressure). The embodiment shown in Figure 10A can utilize stacked membrane assemblies formed of dry fibrillated polymer membranes 1020 each containing inorganic particles. Test Method Method for determining membrane moisture content

[0138] The moisture content of the membranes was determined according to the guidelines set forth in ASTM standard E1868-10, Standard Test Method for Loss on Drying by Thermogravimetry. All data was collected using a Mettler Toledo HB43 Moisture Analyzer (Mettler-Toledo GmbH, CH-8606 Greifensee, Switzerland). The membrane area was consistent at 109.5 cm. 2 Six 2 cm diameter circles were die cut using a die cutter. For wet samples, each circle was gently placed on a low-lint wipe for a few seconds to remove surface moisture before introducing it into the instrument for measurement. Each sample was heated at 110°C for a total of 2 hours. Values ​​were reported as loss on drying (LOD) in % by weight. Methods for determining membrane air permeability, thickness, and density

[0139] To determine the density and thickness of each film sample, a 1-inch (5.3 cm) diameter sample was taken from each film. 2 Circles of 1 / 4" x 1 / 4" were punched out from the sample. The samples were weighed, and then the thickness was measured using a Mitutoyo Litematic VL-50 thickness gauge (Mitutoyo America Corporation, Aurora, IL). The membranes were characterized for air permeability using a Gurley Model 4340 automatic densometer (Gurley Precision Instruments, 514 Fulton Street, Troy, NY 12180). The 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. Methods for determining the dynamic binding and elution behavior of membranes

[0140] To determine the dynamic binding capacity of each membrane sample, a 25 mm diameter (4.9 cm) sample was taken from each membrane. 2 Two circles of 1000µm were die cut. These samples were stacked and held in place with a filter clamp, allowing for controlled flow of the solution through the membrane.

[0141] Two 25 mm disks of membrane sample were placed between a support screen (VWR® Support Screen, stainless steel, 89428-948) and a glass funnel (VWR® Glass Funnel, 15 ml, 89428-938) and placed in a 125 ml graduated filter flask assembly (VWR® Filter Flask Graduated, 125 ml, 89428-978).

[0142] A sample vial (VWR® Sample Vial, clear borosilicate glass, screw thread, 20 ml, 66009-567) was placed into a 125 ml filter flask (VWR® Graduated Filter Flask, 125 ml, 89428-978) with the outlet of the glass base (VWR® Glass Base for Stainless Steel Support, 89428-942) positioned within the sample vial. A support screen (VWR® Support Screen, stainless steel, 89428-948) was placed on the glass base. The membrane sample was placed between the support screen and a glass funnel (VWR® Glass Funnel, 15 ml, 89428-938) and held in place with an aluminum clamp (VWR® Aluminum Clamp, 89428-944).

[0143] A single cycle of binding and elution to determine the dynamic binding and elution performance of the membrane was carried out using the following protocol: The solutions listed in Table A were applied to a glass funnel in the volumes and method order listed in Table B. The eluents obtained at each method step were collected in dedicated sample vials (VWR® Sample Vials, clear borosilicate glass, screw thread, 20 ml, 66009-567) as shown in Table B. The eluent sample vials were changed after each method step. Each solution flowed through the membrane under gravity. [Table 1] [Table 2]

[0144] The concentration of mAb in each elution sample was calculated by measuring absorbance at a wavelength of 280 nm using a Hitachi U-2900 spectrophotometer (Hitachi High-Tech America, 10 North Martingale Road, Suite 500, Schaumburg, IL 60173-2295).

[0145] The concentrations of mAb in the target analyte feed and in the flow-through and wash samples were calculated by measuring absorbance at a wavelength of 280 nm using a Hitachi U-2900 spectrophotometer (Hitachi High-Tech America, 10 North Martingale Road, Suite 500, Schaumburg, IL 60173-2295).

[0146] The mAb concentration of the sample was then calculated using Beer's law (Eq-01). Eq-01 A=e×L×C where: A=absorbance e=1.47mLg -1 cm -1 (Extinction coefficient of the target analyte, trastuzumab biosimilar) L = sample path length C=solution concentration Freeze-drying of membranes

[0147] To dry each Protein A-immobilized membrane, the membrane was hung in place and thoroughly rinsed with deionized (DI) water to ensure that most of the solvent evaporated was water. After the final water soak, each membrane was then frozen at -40°C for 24 hours in a Revco DXF-40040A ultralow temperature freezer (Thermo Fisher Scientific, 81 Wyman Street, Waltham, MA 02451). Each frozen membrane was then transferred to a VirTis 25L Genesis SQ Super ES 55 freeze-drying unit (SP Industries, 935 Mearns Road, Warminster, PA 18974), where both the refrigerator and shelf were frozen before the cycle began. The membrane was then dried after the initial shelf (≤-35°C) and vacuum (≤100 mT) conditions were achieved according to the cycle program settings, following the steps shown in Table C. [Table 3]

[0148] After completing the freeze-drying cycle, each dry membrane was characterized for moisture content according to the Method for Determining Moisture Content of Membranes. Ethylene oxide (EO) treatment of membranes

[0149] Each dry membrane that had been subjected to ethylene oxide (EO) treatment was dried according to the membrane freeze-drying method, then cut and placed in a sterilization pouch (part number PG-7514, plastcareusa, Canoga Park, CA).

[0150] Several membranes were subjected to EO treatment using a 3M Steri-Vac™ Sterilizer GS8X-1D (3M Health Care, St. Paul, MN) under treatment conditions representative of an EO sterilization cycle. In the EO treatment cycle, 170 grams of EO was injected into the sterilizer containing the membranes using a Steri-Gas™ EO gas cartridge (part number 8-170, 3M Health Care, St. Paul, MN), and the sterilizer was held at a temperature of 40°C for 3 hours. Details of this cycle can be found in Table D. [Table 4] Method for determining dynamic binding capacity at 10% breakthrough of device articles

[0151] The chromatography device was inserted into the flow path of an AKTA™ Pure (Cytiva, Marlborough, Massachusetts) liquid chromatography system, and a single cycle consisting of the following protocol was performed: Table E lists the solutions used, and Table F lists the protocol steps for determining the dynamic binding capacity at 10% breakthrough. [Table 5] [Table 6] Method for determining the liquid permeability of an article of equipment

[0152] The liquid permeability of the chromatographic devices was determined using Darcy's law. Individual devices were characterized for packing cross-sectional area and packing length. Solution A was used as the liquid and characterized for viscosity. The pressure drop across the column as a function of liquid flux was measured with an AKTA™ Pure liquid chromatography system (Cytiva, Marlborough, MA). Incipient Wetness of Dry Equipment Articles

[0153] The dry device article was wetted prior to characterization testing. The chromatography device was inserted into the flow path of an AKTA Pure liquid chromatography system, and a buffer solution of 50 mM sodium phosphate, 150 mM sodium chloride, pH 7.4 was pumped through the device, first in the upstream direction, then in the downstream direction, according to the flow rates and volumes in Table G below. [Table 7] Method for determining moisture content of an article of equipment

[0154] The device article was weighed after assembly to obtain the dry mass of the device. The device article was then reweighed after initial wetting to collect permeability and DBC data. The wet mass of the device represents the device when filled with the storage solution specified in Table E. Ethylene oxide (EO) treatment of equipment articles

[0155] Each dry device item that underwent ethylene oxide (EO) sterilization was placed in a sterilization pouch (part number PG-7514, plastcareusa, Canoga Park, CA).

[0156] Several devices were subjected to EO treatment using a 3M Steri-Vac™ Sterilizer GS8X-1D (3M Health Care, St. Paul, MN) under treatment conditions representative of an EO sterilization cycle. The EO treatment cycle involved injecting 170 grams of EO into the sterilizer containing the devices using a Steri-Gas™ EO gas cartridge (part number 8-170, 3M Health Care, St. Paul, MN), and holding the sterilizer at a temperature of 40°C for 3 hours. Details of this cycle can be found in Table H. [Table 8] Method for determining elution volume from 100mAU to 100mAU of an article of equipment

[0157] The wetted chromatography device was inserted into the flow path of an AKTA™ Pilot 600 (Cytiva, Marlborough, Massachusetts) liquid chromatography system, and a single cycle consisting of the following protocol was performed: Table I lists the solutions used, and Table J lists the steps of the purification protocol used. Chromatograms of UV absorbance at 280 nm generated according to the purification protocol in Table J were used to determine the elution volumes from 100 mAU to 100 mAU. [Table 9] [Table 10]

[0158] The UV absorbance chromatogram at 280 nm generated following the purification protocol in Table J was plotted against the cumulative volume passed through the device and used to determine the elution volume of the device article from 100 mAU to 100 mAU using Eq-02.

[0159] Eq-02 EV 100~100 =CV 100終了 -CV 100開始 where: EV 100~100 = elution volume from 100mAU to 100mAU, CV 100開始 = cumulative volume of buffer and CHO cell harvest passed through the device up to the start of elution (column volume CV), defined as the point at which the UV absorbance at 280 nm exceeds 100 mAU when the device is loaded with elution buffer according to step 4 of Table J. CV 100終了 = cumulative volume of buffer and CHO cell harvest passed through the device by the end of elution (column volume, CV), defined as the point at which the UV absorbance at 280 nm falls below 100 mAU when the device is loaded with elution buffer according to step 4 of Table J. Example

[0160] Membrane A was a porous polytetrafluoroethylene (ePTFE) membrane having 15 weight percent ePTFE and 85 weight percent porous spherical silica particles. Membrane B was a porous ePTFE membrane having 15 weight percent PTFE and 85 weight percent porous irregularly shaped silica particles. Membranes A and B were identical to those described in U.S. Patent Application Publication No. 2023 / 0356109 by Clinger et al. Table K lists some of the physical properties of the two porous ePTFE membranes. [Table 11]

[0161] The membrane was then processed to covalently bond (immobilize) Protein A to the ePTFE membrane in a manner representative of those skilled in the art and further described in U.S. Patent No. 10,525,376 to McManaway et al. and U.S. Patent No. 10,526,367 to McManaway et al., to produce Protein A-immobilized Membranes C and D, respectively.

[0162] The Protein A-immobilized membranes are freeze-dried according to the membrane freeze-drying method described above to produce dry Protein A-immobilized membranes E and F, respectively.

[0163] Sections of dry immobilized membranes E and F are subjected to ethylene oxide (EO) treatment according to the membrane ethylene oxide treatment method described above to produce dry Protein A immobilized ethylene oxide treated membranes G and H, respectively.

[0164] The membranes whose manufacture is 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 dry membrane. The performance of each of these affinity chromatography membranes is shown in Table L. [Table 12]

[0165] Non-dry membranes C and D, the preparation of which is described above, were used to fabricate non-dry reference affinity chromatography devices in stacked membrane and parallel-branched formats. Dry membranes E through H, the preparation of which is described above, were used to fabricate dry affinity chromatography devices in stacked membrane, spiral-wound, and parallel-branched formats. The membranes and conditions associated with each non-dry and dry affinity chromatography device are summarized in Table M. The dry affinity chromatography devices were wetted with 50 mL of sodium phosphate and 150 mM sodium chloride buffer using the method described above and then tested to evaluate their liquid permeability and dynamic binding capacity at a 20-second residence time using the protocol described in the test methods. The performance of these devices is reported in Table N. [Table 13] [Table 14]

[0166] Device T was further characterized for elution volume performance using a method for determining the elution volume of a device article from 100 mAU to 100 mAU. The performance of Device T is shown in Table O. A chromatogram of UV absorbance at 280 nm associated with the characterization of Device T using a method for determining the elution volume of a device article from 100 mAU to 100 mAU is shown in Figure 9. [Table 15]

[0167] 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 in the embodiments without departing from the scope of the present disclosure. Therefore, it is intended that the embodiments include the modifications and variations of the present invention provided that they come within the scope of the appended claims and their equivalents.

Claims

1. 1. A composite membrane for an affinity chromatography device, the composite membrane comprising: a dry fibrillated polymer membrane containing inorganic particles therein, wherein at least one of the inorganic particles and the dry fibrillated polymer membrane has covalently attached thereto an affinity ligand that reversibly binds to a target molecule; A composite membrane, wherein the dry fibrillated polymer membrane has a moisture content of 60% or less by weight of the dry fibrillated membrane.

2. The composite membrane of claim 1 , wherein the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

3. 3. The composite membrane of claim 1 or 2, wherein the inorganic particles have a spherical shape and a nominal particle size of about 5 micrometers to about 20 micrometers.

4. 4. The composite membrane of claim 1, wherein the inorganic particles have a particle size distribution D90 / D10 of 3 or less.

5. 5. The composite membrane of claim 1, 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. 6. The composite membrane of claim 1, wherein the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, an oligonucleotide, and a combination thereof.

7. The composite membrane of claim 1 , wherein the target molecule comprises a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

8. 8. The composite membrane of claim 1, wherein the dry fibrillated polymer membrane has a wound configuration.

9. 9. The composite membrane of claim 1, wherein the dry fibrillated polymer membrane has a stack configuration.

10. 10. The composite membrane of claim 1, wherein the dry fibrillated polymer membrane is sterilized by ethylene oxide, gamma irradiation, or X-ray irradiation.

11. 1. An affinity chromatography device, comprising: a fluid inlet; a fluid outlet in fluid communication with the fluid inlet; a dry-fibrillated polymer membrane disposed between the fluid inlet and the fluid outlet, the dry-fibrillated polymer membrane having at least one inorganic particle therein; Including, the dry fibrillated polymer membrane has a moisture content of 60% or less by weight of the dry fibrillated polymer membrane before drying; or The affinity chromatography device has a water content of 60% by mass or less of the affinity chromatography device. At least one of the following applies, and A device wherein at least one of the dry fibrillated polymer membrane and the inorganic particles is covalently bound to an affinity ligand that reversibly binds to a target molecule.

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

13. 13. The apparatus of claim 11 or 12, wherein two or more of the dry fibrillated membranes are in the form of a dry stack membrane assembly disposed between the fluid inlet and the fluid outlet.

14. 13. The device of claim 11 or 12, wherein the dry fibrillated membrane has a wound membrane configuration comprising a dry fibrillated polymer wound around a core.

15. The device of claim 11 , wherein the target molecule comprises a protein, an antibody, a viral vector, a nucleic acid, or a combination thereof.

16. 16. The device of claim 11, wherein the inorganic particles are selected from spherical particles, non-spherical particles, and combinations thereof.

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

18. 18. The device of any one of claims 11 to 17, wherein the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, and combinations thereof.

19. 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 has been treated with ethylene oxide, gamma irradiation, or X-ray irradiation.

20. 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 affinity chromatography device has a capacity of about 100 (×10 －１２ cm ２ ) ~ approx. 2000 (×10 －１２ cm ２ 21. The device of claim 11, configured to achieve a water permeability of 0.1% to 1.2%.

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

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

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

26. 26. The manifold of claim 25, wherein the manifold is contained within a single housing.

27. 24. An apparatus 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 described in any one of claims 11 to 23.

28. 28. The apparatus of claim 27, wherein the first manifold and the second manifold are contained within a single housing.

29. 1. A method for isolating a target molecule, the method comprising: The entrance and The exit and a dry fibrillated polymer membrane having a moisture content of 60% by mass or less of the dry fibrillated polymer membrane; providing a chromatography device comprising: wherein the dry fibrillated polymer membrane is disposed between the inlet and the outlet and comprises at least one inorganic particle therein; wetting the dry fibrillated polymer membrane to form a wet polymer membrane; adding an aqueous mixture containing a target molecule to the inlet; passing the aqueous mixture through the wet fibrillating polymer membrane, thereby binding the target molecule to the affinity ligand; binding the target molecule to the affinity ligand, wherein the affinity ligand is present on at least one of the dry fibrillated polymer membrane and the inorganic particles; A method comprising:

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

31. 31. The method of claim 30, wherein the treatment is carried out according to the Ethylene Oxide (EO) Treatment Method for Membranes described in the Test Methods section.

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

33. 30. 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. 30. The method of claim 29, comprising treating a non-dry fibrillating membrane to covalently bond affinity ligands to one or both of the non-dry fibrillating polymeric membrane and the inorganic particles.

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

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

37. 30. The method of claim 29, comprising characterizing the water content of the chromatography device.

38. 30. The method of claim 29, comprising drying a non-dry fibrillated membrane by lyophilization to form said dry fibrillated polymeric membrane.

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

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

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

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

43. 43. The method of claim 42, wherein the initial wetting step is performed according to the Dry Equipment Article Incipient Wetness Method described in the Test Methods section.

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

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

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

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

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

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

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

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

52. 1. An affinity chromatography device, comprising: a housing member; an inlet for allowing fluid to enter the housing member; an outlet for permitting fluid to exit the housing member and in fluid communication with the inlet; a stack membrane assembly disposed within the housing member between the fluid inlet and the fluid outlet; the stack membrane assembly comprising: two or more dry fibrillated polymer membranes in a stacked configuration, each of said dry fibrillated polymer membranes comprising inorganic particles therein; the dry fibrillated polymer membrane has a moisture content of 60% by mass or less of the dry fibrillated polymer membrane; A device wherein at least one of the dry fibrillated polymer membrane and the inorganic particles is covalently bound to an affinity ligand that reversibly binds to a target molecule.

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

55. 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 micrometers to about 20 micrometers.

56. 56. An apparatus according to any one of claims 52 to 55, wherein the particle size distribution has a D90 / D10 of 3 or less.

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

58. 58. The device of any one of claims 52 to 57, wherein the affinity ligand is selected from protein A, protein G, protein L, a human Fc receptor protein, an antibody, a polysaccharide, an oligosaccharide, an oligonucleotide, and combinations thereof.

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

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

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