Radial flow separation device

Abstract

This disclosure provides a radial flow separation device comprising a capsule body, a lid that attaches to the capsule body, and a separation media housed within the capsule body. Fluid flows through either of a first connector or a second connector and then travels through the separation media in a direction radial to a direction in which flow passes through the second connector. The separation device is particularly suitable for purification and separation of target components, including, biological products, intended for use as biotherapeutics.

Description

TITLE: RADIAL FLOW SEPARATION DEVICECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of Great Britain Provisional Application Ser. No.2311253.5 filed on July 21, 2023 and titled “RADIAL FLOW SEPARATION DEVICE”, hereby incorporated by reference in its entirety.TECHNICAL FIELD

[0002] The present disclosure relates generally to a radial flow separation device and / or corresponding method(s) of use / manufacture. The radial flow separation device has applications in industries where there is a desire to separate and capture target components such as biomolecule, viruses, cells and other materials. This disclosure is particularly pertinent to bioseparations.BACKGROUND

[0003] The background description provided herein gives context for the present disclosure. Work of the presently named inventors, as well as aspects of the description that may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art.

[0004] In medicine, biotherapeutics, vaccines and other treatments are increasingly reliant on the separation of particular target components from a number of other non-target components. Target components can vary in size and properties, including even fragments of molecules, viruses and nucleic acid materials. The ability to capture specific components efficiently requires separation devices and methods that can handle high volumes of fluid at increasingly fast flow rates. The media utilized in bioseparations can vary from membranes to beads (including resins) and other materials. In order to effectively capture targeted components, it is necessary to have separation devices that house the media and provide desirable fluid dynamics including even fluid paths, laminar flow, and desired pressure. Additionally, the ability to increase the surface area per volume of the media capable of capturing targeted components is important to improving the efficiency of the bioseparation process.

[0005] Conventional purification methodologies, such as column chromatography separations and certain membrane-based technologies, are limited in terms of binding capacity and processing time. These limitations are primarily due to slow diffusion rates of relatively large biomolecules, which limits the ability of the target component to access available binding sitesdeep within the separation media. In addition, these systems can be extremely large and require excessive amounts of separation media.

[0006] Various media and configurations have been utilized over the years including beads (see, e.g., WO 1996 / 009116) and membrane technologies in different configurations, including, pleated and rolled (see, e.g., U.S. 2013 / 0092620) and layered (see, e.g., U.S. 9,120,037). However, separation devices with increased binding capacity, maximized flow rates of the mobile phase, and improved usage and pressure across the entire separation media have lagged behind the corresponding improvements in ligand and binding chemistries. Thus, while various configurations have been achieved for different separation media (such as membranes and resins), the existing device configurations still fail to achieve the potential of the separation media themselves.

[0007] Radial-style filtering devices have been utilized more extensively in other technology areas, for example as described in U.S. Patent No. 7,645,312 (owned by 3M), where a radial filter device is described for separating contaminants by passing a fluid through porous filter so as capture via size exclusion contaminants particles while permitting the fluid to pass therethrough. However, such filters are highly distinct in their design and function and frequently incorporate a spiral-wound membrane.

[0008] Thus, there remains a need for improved separation devices, including, devices that have improved binding capacity on a mass basis, improved binding capacity on a volume basis, improved flux, reduced dead volume, substantially uniform pressure across the separation media, improved flow speeds while still achieving separation, and / or achievement of substantially laminar flow.SUMMARY

[0009] The following objects, features, advantages, aspects, and / or embodiments, are not exhaustive and do not limit the overall disclosure. No single embodiment need provide each and every object, feature, or advantage. Any of the objects, features, advantages, aspects, and / or embodiments disclosed herein can be integrated with one another, either in full or in part.

[0010] It is a primary object, feature, and / or advantage of the present disclosure to improve on or overcome the deficiencies in the art.

[0011] For example, a radial flow separation device is provided which beneficially passes flow through the separation media in the direction radial to the first port direction. The deviceincludes a lid, a capsule body, and a separation media, with the separation media sealed inside the capsule body with elastomeric encapsulation members. All other features of the device as described herein are optional, non-essential features of the invention, though several of these optional features are novel and inventive in their own right.

[0012] A preferred embodiment disclosed herein is a separation device comprising a capsule body, a separation media assembly located at least partially within the capsule body, the separation media assembly comprising a separation media sealed (preferably hermetically sealed) within the capsule body, a first encapsulation member, and a second encapsulation member, wherein the separation media comprises one or more surface functionalizations, the separation media is pleated or rolled, and the separation media forms a center void, a core positioned in the center void of the separation media, a lid, a first port proximate to the capsule body, and a second port proximate to the lid, characterised in that fluid can flow between the first port and the second port and can pass through the separation media in a direction radial to a direction in which fluid passes between the first port and / or second port. Related methods of controlling fluid through the separation device and methods of separating one or more target components are also disclosed herein.

[0013] A preferred embodiment disclosed herein is a system for separating one or more target molecules, the system comprising an upstream flow line, a pump, a separation device, a downstream flow line, optionally one or more instruments and / or sensors, a process controller and a fraction collector. The separation device comprises a capsule body having a second port, a separation media assembly located at least partially within the capsule body and having a center void, a core positioned in the center void of the separation media, and a lid having a first port; wherein the separation media assembly comprises a separation media sealed (preferably hermetically sealed) within the capsule body, a first encapsulation member, and a second encapsulation member; wherein the separation media comprises one or more surface functionalizations; and wherein the separation device is characterised in that fluid that flows between the first port and second port and passes through the separation media in a direction radial to a direction in which fluid passes through the first port and / or second port. The upstream flow line is in fluid communication with the first port, and the downstream flow line is in fluid communication with the second port. Related methods of controlling fluid through the system and methods of separating one or more target components are also disclosed herein.

[0014] It is to be appreciated that the aforementioned features each contribute toward a separation device that is safe to use, cost effective, and durable. Together, they synergistically and surprisingly outperform known devices in the art, as outlined in the EXAMPLES section provided herein.

[0015] The separation device disclosed herein can be used in a wide variety of applications. For example, the separation device can be used once for a purification application or re-used multiple times. Additionally, but non-limitingly, the separation device can be employed in rapid cycling processes to beneficially provide ultrafast purification with a relatively small separation form factor. Additionally, but non-limitingly, the separation device can be used in flexible operations on a large scale.

[0016] Methods can be practiced which facilitate use, manufacture, assembly, maintenance, and repair of separation device(s) which accomplish some or all of the previously stated features and / or objectives.

[0017] The separation device(s) described herein can be incorporated into systems or kits which have some and / or all of the features described above and / or accomplish some or all of the previously stated objectives. Multiple separation devices can be incorporated into systems or kits by connecting separation devices together in series or in parallel.

[0018] These and / or other objects, features, advantages, aspects, and / or embodiments will become apparent to those skilled in the art after reviewing the following brief and detailed descriptions of the drawings. The present disclosure encompasses (a) combinations of disclosed aspects and / or embodiments and / or (b) reasonable modifications not shown or described.

[0019] In a first aspect of the invention there is provided a separation device comprising: a capsule body; a separation media assembly located at least partially within the capsule body; the separation media assembly comprising a separation media sealed (preferably hermetically sealed) within the capsule body and a first encapsulation member and a second encapsulation member; wherein the separation media is pleated or rolled, and the separation media assembly includes a center void; a core positioned in the center void of the separation media; a lid; a first connector proximate the lid providinging a first port for fluid to pass through; a second connector proximate the capsule body providing a second port for fluid to pass through; and characterised in that fluid can flow between the first connector and the secondconnector and can pass through the separation media in a direction radial to a direction in which fluid passes through the second connector.

[0020] The separation media may comprise one or more surface functionalizations.

[0021] The lid may be friction-fitted, welded, screwed, clamped, Luer-locked, glued, adhered or otherwise secured together and sealed to the capsule body.

[0022] The lid may further comprise a primary interface that hygienically couples a geometry of the first connector with an inside region of the separation media assembly.

[0023] The primary interface may comprise a radial static seal established with a connector formed into the lid and directly coupled to a sealing face on an underside of the lid; preferably wherein the connector comprises any of a Luer, a Luer-lock, a tri-clamp connection, a ferrule, and / or a threaded fitting.

[0024] A thread of the threaded fitting comprises any of a Unified national fine (UNF) thread, a G-thread, a national pipe thread (NPT), or a metric thread, or any other threading type.

[0025] The first encapsulation member may be made of an elastomeric material and is sealed (preferably hermetically sealed) to the separation media.

[0026] The elastomeric material may be a silicone elastomer, a polyurethane elastomer, or a combination thereof.

[0027] The second connector may be centrally located on the capsule body.

[0028] The first connector may be centrally located on the lid.

[0029] Alternatively, the first connector and the second connector may extend in a same direction from the separation device in a common plane.

[0030] The primary interface may comprise a flange static seal established by a connector that is part of an insert that interfaces with the lid.

[0031] The connector may comprise any of a Luer, a Luer-lock, a tri-clamp connection, a ferrule, and / or a threaded fitting.

[0032] A thread of the threaded fitting may comprise any of a Unified national fine (UNF) thread, a G-thread, a national pipe thread (NPT), and / or a metric thread, or any other threading type.

[0033] The insert may compress a sealing ring that is integral to a first encapsulation member between an insert flange and the lid.

[0034] The separation device may further comprise a retaining feature that maintains compression between a retaining component and the lid.

[0035] The retaining component may be threaded, grooved, or moulded in-place.

[0036] The first encapsulation member may have an extended portion.

[0037] The second encapsulation member may be made of an elastomeric material and is sealed to the separation media.

[0038] The elastomeric material may be a silicone elastomer, a polyurethane elastomer, or a combination thereof.

[0039] The lid may further comprise a secondary interface with the separation media assembly that provides a hygienic seal between the lid and the capsule body, and wherein the secondary interface comprises a ridge that interacts with the first encapsulation member.

[0040] The capsule body may comprise a primary interface that hygienically couples the capsule body with an outside region of the separation media assembly.

[0041] The first connector may be parallel with the second connector and the flow is passed through the separation media in a direction radial to a direction in which flow passes through the second connector.

[0042] The separation device may further comprise a connector that is formed into the capsule body.

[0043] An interior radial wall of the capsule body and an exterior surface of the separation media may provide an outer channel that tapers.

[0044] An interior surface of the separation media of the capsule body and the core may provide an inner channel that tapers.

[0045] The outer channel and the inner channel may taper at a slope inverse to each other.

[0046] The interior radial wall may interface with a second encapsulation member such that the flow is permitted with a controlled resistance.

[0047] The first encapsulation member and second encapsulation member may be made of the same elastomeric material.

[0048] The core may be made of the same elastomeric material.

[0049] The separation media may comprise a join that is thermally bonded or encapsulated.

[0050] The capsule body may comprise internal ridges.

[0051] The core maintains an equidistant flow-path reducing void volume inside the separation media.

[0052] The core may be hygienically coupled to a geometry of the lid and tapered.

[0053] The core may further comprise one or more fins and / or vents.

[0054] A fluid may enter the separation device via a first connector and the fluid is directed by a diverter to an interior surface of the separation media.

[0055] The separation media may comprise a hydrogel, one or more membranes, a monolith, or a matrix.

[0056] The separation media may comprise one or more fibrous membranes or a cast membrane.

[0057] The fibrous membrane may comprise microfibers, nanofibers, or a mixture thereof.

[0058] The fibrous membrane may comprise an electrospun membrane.

[0059] The separation media may comprise multiple layers, wherein the fibrous membrane comprises a primary functional layer attached to a secondary inert layer.

[0060] The separation device may further include a conduit attached to the lid and the first connector, wherein the first port extends through the conduit, wherein the first connector is proximate the lid via the conduit, wherein the conduit redirects flow substantially 180- degrees with respect to flow that passes through the second connector.

[0061] The separation device may further be configured such that the lid further comprises a side aperture through which the conduit passes.

[0062] The separation device may further include a bleed-valve disposed on the capsule body.

[0063] The separation device may further include the bleed-valve is adjacent to the second connector.

[0064] The separation device may further include a first frit disposed between the lid and the second encapsulation member.

[0065] The separation device may further include a second frit disposed between the core and the capsule body.

[0066] The separation media may comprise a polymer such as collagen, chitosan, agarose, agarose acetate, cellulose, cellulose acetate, crosslinked cellulose, derivatized cellulose, regenerated cellulose, polyacrylic acid, polymethacrylic acid, polyacrylonitrile, a polyethylene oxide, a polyimide, a polyamide, polystyrene, a polysulfone, polycaprolactone, or a copolymer or mixture thereof.

[0067] The one or more surface functionalizations may be in a density on the separation media of from about 1 mg / g to about 999 mg / g.

[0068] The surface functionalized separation media may have a dynamic binding capacity on a volume basis of between about 1 pg / mL of the separation media and about 400 mg / mL of the separation media.

[0069] The one or more surface functionalizations may comprise an ion exchange group, a polar group, a hydrophobic group, an affinity ligand, a mixed mode ligand, or a combination thereof.

[0070] The one or more surface functionalizations may comprise an anion exchange group, a cation exchange group, or a mixture thereof.

[0071] The ion exchange group may comprise one or more of a tertiary amine, a quaternary amine, a quaternary ammonium, 6-chloranyl-3-[(2-pentyl-2,3-dihydro-l,3-thiazol-4- yl)methyl]quinazolin-4-one, diethyl-(2-hydroxy-propyl) aminoethyl, diethylaminoethyl, trimethylaminoethyl, — N+(CH3)3, — N+(C2H5)H, — CH2CH2N+(CH3)3, — O— CH2CH2— N+(CH3)3, — CH2CH2N+(C2H5)H, — CH2CH2N+(C2H5)2(CH2CH(OH)CH3), — CH2CH2N+(CH3)2H, a carboxylate, a sulphonate, a phosphonate, — CH2COO“, — O- CH2COO“, — CH2OCH2COO“, — SO3“, — CH2CH2CH2SO3’, — CH2CH2SO3’, — P(OH)2O“, or a mixture thereof.

[0072] The one or more surface functionalizations may comprise a hydrophobic group.

[0073] The hydrophobic group may comprise a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, a hexyl group, an octyl group, a decyl group, a phenyl group, or a mixture thereof.

[0074] The one or more surface functionalizations may comprise an affinity ligand.

[0075] The affinity ligand may comprise a monoclonal antibody, a polyclonal antibody, an antibody fragment, a bacterial immunoglobulin binding protein, a chemical ligand, a dye, an enzyme inhibitor, histidine, an immobilized metal ion, a nucleic acid, an oligonucleotide, a lectin, a protein, an oligopeptide, a polysaccharide, an oligosaccharide, a sugar, a peptide, a polypeptide, an antigen, an aptamer, an affimer, an affibody, a small molecule biomimetic ligand, a small organic compound, a synthetic affinity ligand, a triazine ligand or a mixture thereof.

[0076] The Affinity ligand may be Protein A, Protein G, or Protein L.

[0077] The affinity ligand may comprise IgG, IgM, a monoclonal antibody, a camelid antibody, an antibody fragment, Fab, Fv, or a mixture thereof.

[0078] The one or more surface functionalizations may comprise a mixed mode ligand.

[0079] The mixed mode ligand may comprise at least two of an anion exchange group, a cation exchange group, a polar group, a hydrophobic group, and / or a hydrogen bonding group.

[0080] The mixed mode ligand may comprise an N-benzyl methyl ethanolamine group, an N- benzoyl-homocysteine group or mixture thereof.

[0081] In a second aspect of the invention there is provided a method of controlling flow through the separation device of the first aspect, comprising: flowing a fluid through the lid at the first connector; passing the fluid through the separation media; wherein the fluid passes through the separation media in a direction radial to a direction in which fluid passes through the second connector; and passing the fluid through the second connector.

[0082] The core and / or a diverter may direct the flow to an inner channel which tapers.

[0083] The fluid may exit the separation media into an outer channel which tapers.

[0084] The outer channel and the inner channel may taper inverse to each other.

[0085] In a third aspect of the invention there is provided a method of controlling flow through the separation device of the first aspect, comprising: flowing a fluid through the capsule body at the second connector; passing the fluid through separation media; wherein the fluid passes through the separation media in a direction radial to a direction in which flow passes through the second connector; and passing the fluid through the first connector.

[0086] The core and / or a diverter may direct the flow to an inner channel which tapers.

[0087] The fluid may exit the separation media into an outer channel which tapers.

[0088] The outer channel and the inner channel may taper inverse to each other.

[0089] The fluid flow may be laminar and an equivalent residence-time within the device is achieved by provisioning equidistant paths through the separation device.

[0090] The method may be further characterised by reversing the flow.

[0091] In a fourth aspect of the invention there is provided a method of purifying a target component comprising: flowing a fluid through the first connector or second connector of the separation device according to the first aspect; passing the fluid through separation media; wherein the fluid passes through the separation media in a direction radial to a direction in which flow passes through the first connector; separating the biomaterial from the fluid; and passing the fluid through the second connector.The target component may comprise an amino acid, a biomolecule or fragment thereof, a peptide, an affimer, a protein, an enzyme, a glycoprotein, an LNP, a lipopolysaccharide, an antibody and / or a fragment thereof, a nucleic acid, an organic polymer, a virus, a VLP, an extracellular vesicle, an exosome, a bacterium, a cell, a cell-related structure, or a mixture thereof.The antibody may comprise a monoclonal antibody, a single-chain antibody, a bi-specific antibody, a multi-specific antibody, an antibody conjugate, an antibody fusion protein, a drug-antibody conjugate, an antibody fragment, a Fab fragment, a Fv fragment, a gamma globulin (IgG), an IVIG, an IgM, an IgA, an IgE, a hyperimmune gamma globulin, an isoagglutinin, or a combination thereof.

[0092] The virus may comprise adeno-associated virus, lentivirus, or mixture thereof.

[0093] The fluid flow may be performed at a pressure of about 1 bar or less.

[0094] The fluid flow may be performed at a pressure of about 0.2 bar or less.

[0095] The residence time may be less than about 1 minute.

[0096] The residence time may be about 15 seconds or less.

[0097] In a fifth aspect of the invention there is provided a system for separating one or more target molecules comprising: a separation device comprising a capsule body having a second connector; a separation media assembly located at least partially within the capsule body, the separation media assembly comprising a separation media sealed (preferably hermetically sealed) within the capsule body and a first encapsulation member and a second encapsulation member, wherein the separation media comprises one or more surface functionalizations and the separation media assembly includes a center void; a core positioned in the center void of the separation media; and a lid having a first connector; characterised in that fluid that flowsbetween the first connector and second connector and passes through the separation media in a direction radial to a direction in which fluid passes through the first connector and / or second connector; an upstream flow line in fluid communication with the first connector, and a downstream flow line is in fluid communication with the second connector.

[0098] The system further comprising one or more instruments and / or sensors in fluid communication with the downstream flow line.

[0099] The system, wherein the one or more instruments and / or sensors may be, but are not limited to, a chromatography work station, a fast protein liquid chromatography system (“FPLC”), an Akta system, a process chromatography skid, a membrane chromatography skid, a high performance liquid chromatography instrument (“HPLC”), an ultra-performance liquid chromatography instrument (“UPLC”), a next generation chromatograph (“NGC”), a mass chromatograph (“MS”), a gas chromatograph (“GC”), a colorimeter, a mass spectrometer, a fluorometer, a photometer, a spectrometer, a spectrophotometer, a X-ray photoelectron spectrometer (“XPS”), a sequencing instrument, and a next generation sequencing (“NGS”) sequencing instrument, a UV detector, a visible light detector, a pH sensor, a pressure sensor, a temperature sensor, a conductivity sensor, or combination thereof.

[0100] Alternatively, the system might comprise a pumping system such as a peristaltic pump, a diaphragm pump, a syringe pump, a positive displacement pump, a quattroflow pump, a centrifugal pump and a lobe pump connected in series and in fluid communication with a separation device and one or more instruments or sensors including, but not limited to, a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector or a diode array detector, or any combination thereof, any or all of which may optionally be connected to a recording device and / or a controller device. Thus, the system need not be limited to a chromatography work station, an FPLC, an Akta system, a process chromatography skid, a membrane chromatography skid, or other traditional chromatography systems.

[0101] The system further comprising a fraction collector in fluid communication with the downstream flow line.BRIEF DESCRIPTION OF THE FIGURES

[0102] Several embodiments in which the present disclosure can be practiced are illustrated and described in detail, wherein like reference characters represent like components throughoutthe several views. The figures are presented for exemplary purposes and may not be to scale unless otherwise indicated.

[0103] Figure 1 shows an exploded, perspective view of a separation device where flow is passed through the separation media unit in a direction radial to the direction of flow upon entering the first connector.

[0104] Figure 2A shows a detailed, underside of a lid 100 of the separation device of Figure 1

[0105] Figure 2B shows a detailed, sectional, and cut-out view from Figure 2A of the lid 100 of the separation device of Figure 2A.

[0106] Figure 3A shows a detailed, internal perspective view of a capsule body 200 of the separation device of Figure 1.

[0107] Figure 3B shows a detailed, sectional, and axial view of the capsule body 200 of the separation device of Figure 3A.

[0108] Figure 4 shows a detailed, sectional, and axial view of a first embodiment of a separation device that incorporates a radial static seal as a primary interface, designed to hygienically couple the first connector geometry with the inside region of the separation media.

[0109] Figure 5 shows a detailed, sectional, and axial view of a second embodiment of a separation device that incorporates a flange static seal as a primary interface, designed to hygienically couple the first connector geometry with the inside region of the separation media.

[0110] Figure 6 shows a detailed, sectional, and axial view of a secondary interface with the separation media that provides the hygienic seal between the lid and capsule body in another embodiment of a separation device.

[0111] Figure 7 shows a sectional and axial view of a primary interface with the separation media, designed to hygienically couple the capsule body with an outside region of the separation media in an embodiment of a separation device.

[0112] Figure 8 shows a detailed, perspective view of a separation media assembly 300 in an embodiment of a separation device.

[0113] Figure 9 shows a detailed, sectional, and front view of an embodiment of the separation device, emphasizing view of a core 600 that is positioned in the center of the separation media.

[0114] Figures 10A-10D show various views of separation media included within embodiments of the separation device. Figure 10A shows a perspective view thereof. Figure 10B captures a detailed, photographic view of a multi-layered separation media 312. Figure10C shows a top plan, geometric view of a separation media 312 having an encapsulated join 314. Figure 10D captures a top hemispheric view thereof.

[0115] Figure HA shows a detailed, sectional, and perspective view of and embodiment of the separation device, showing how fluid enters and travels through the device in a first embodiment of flow path; notably, as this is a sectional view the flow outward through the membrane would be radial.

[0116] Figure 11B is an enlarged view of the lower right comer portion of Figure 11A such that the flow channels 604 on the second encapsulation member 310 are more easily viewed.

[0117] Figure 11C shows a schematic view emphasizing with examples that where laminar flow-lines are drawn, each has an equivalent residence-time within the device.

[0118] Figure 11D shows a detailed perspective view of an embodiment of a separation media assembly 300 such that optional flow channels 604 can be viewed on the second encapsulation member 310.

[0119] Figure HE shows a detailed, sectional, and perspective view of an embodiment of the separation device, showing how fluid enters and travels through the device in a second embodiment of flow path; notably, as this is a sectional view the flow inward through the membrane would be radial, but enters and exits opposite the flow path shown in Figure 11 A. Figure HE also shows an embodiment comprising an optional bleed-valve 450.

[0120] Figure HF shows a detailed, sectional, and perspective view of an embodiment of the separation device, showing how fluid enters and travels through the device in the notably, as this is a sectional view the flow inward through the membrane would be radial, but enters and exits opposite the flow path shown in Figure HA. Figure HF also shows an embodiment comprising optional frits above and below said separation media assembly 300; the frits 800, 850 can be included in any of the earlier embodiments including that shown in Figures HA and HE and used in conjunction with the optional bleed-valve 450.

[0121] Figure 12A shows sectional perspective view of an embodiment separation device 1000

[0122] Figure 12B shows an exploded view of the embodiment separation device 1000 of Figure 12A.

[0123] Figure 12C shows an elevation view of the embodiment separation device 1000 of Figure 12A.

[0124] Figure 12D shows a cross-section view of the embodiment separation device 1000 of Figure 12C along A-A.

[0125] Figure 13A shows sectional perspective view of an embodiment separation device 2000 having the first and second connectors on the same side of the separation device 2000; the separation device includes an optional bleed-valve 2450 and can optionally further comprise frits as shown in Figure 11F.

[0126] Figure 13B shows an exploded view of the embodiment separation device 2000 ofFigure 13A.

[0127] Figure 13C shows an elevation view of the embodiment separation device 2000 ofFigure 13A.

[0128] Figure 13D shows a cross-section view of the embodiment separation device 2000 of Figure 13C along B-B.

[0129] Figure 14A shows a diagram of the separation device 000, 1000, 2000 of Figures 11A, HE, HF, 12A-D, and / or 13A-13D, wherein the second connector 402, 1402, 2402 is connected to a downstream flow line 704 and the first connector 204, 1204, 2204 is connected to an upstream flow line 702.

[0130] Figure 14B shows a diagram of the separation device 000, 1000, 2000 of Figures HA, HE, 11F, 12A-D, and / or 13A-13D, wherein the first connector 204, 1204, 2204 is connected to a downstream flow line 704 and the second connector 402, 1402, 2402 is connected to an upstream flow line 702.

[0131] Figure 14C shows a top perspective view of an embodiment separation device with fluid lines connected to both the first connector and second connector in a separation system. Notably the system configuration shown in Figure 14 could be used for the devices depicted in Figures HA, HE, 11F, and / or 12A-D.

[0132] Figure 15 shows the breakthrough profile (A280) during BSA loading onto a separation device (000) fitted with 1 ml of DEAE functionalized, pleated, fibrous membrane.

[0133] Figure 16 shows a chromatograph of the BSA elution profile for a separation device (000) fitted with 1 ml of DEAE functionalized, pleated, fibrous membrane.

[0134] Figure 17 provides a comparison of the BSA binding capacity and elution profile of a 1 ml membrane volume separation device (000) compared to 3 commercially available 1 ml anion-exchange membrane chromatography units.

[0135] Figure 18 shows elution peaks for separation device (000) (Astrea DEAE) compared to the elution profiles for 2 commercially available anion-exchange membrane chromatography units.

[0136] Figure 19 shows overlayed elution peak profiles for separation device (000) and separation device (1000).

[0137] Figure 20 shows overlayed transition curve profiles for separation device (000), separation device (1000) and separation device (2000).

[0138] An artisan of ordinary skill in the art need not view, within isolated figure(s), the near infinite distinct combinations of features described in the following detailed description to facilitate an understanding of the present disclosure.DETAILED DESCRIPTION

[0139] Disclosed herein is a separation device, demonstrative embodiments of which are depicted as 000, 1000, and 2000 in the figures; the separation device 000, 1000, 2000 comprises a separation media 312, 1312, or 2312 in a radial orientation; the separation media 312, 1312, 2312 is particularly useful for separation of target components and has many benefits over existing separation devices for separation of targeted components. For example, the separation device 000, 1000, 2000 can achieve high fluid flow rates with low residence time while maintaining low pressure to provide highly efficient separation of target components. A rolled membrane can be utilized in the separation device 000, 1000, 2000; although a more preferred embodiment is a pleated design of the separation media 312, 1312, 2312 which provides a large surface area for the fluid to contact when traveling through the separation device 000, 1000, 2000. Regardless of whether rolled or pleated, the separation media 312, 1312, 2312 can comprise one or more layers of membrane and / or inert material.

[0140] Various features and components of the radial flow separation device disclosed herein are common between different sizes and configurations of the separation device. For example, different configurations of the separation device are depicted as 000, 1000, and 2000 within the figures. It should be understood though that the separation media 312 and separation media assembly 300 are common amongst the different configurations depicted as 000, 1000, and 2000; these common features and components are referenced with the same reference characters and sometimes with a prefix to specify reference to a particular drawing. For example, the separation media assembly has reference character 300 and is referred to as 300 in reference to the device depicted as 000, and then correspondingly has referencecharacter 1300 in reference to the figures depicting separation device 1000 and reference character 2300 in reference to the figures depicting separation device 2000. That said, the features shown and described as related to 300 also apply to 1300 and 2300 as this is a technical feature or component common between the different separation device configurations. Further, certain figures depict optional components, which can be incorporated into other configurations.

[0141] The embodiments of this disclosure are not limited to particular target components, chromatography or capture chemistries, or therapeutic fields. It is further to be understood that all terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting in any manner or scope. For example, as used in this specification and the appended claims, the singular forms “a,” “an” and “the” can include plural referents unless the content clearly indicates otherwise.

[0142] Definitions

[0143] All units, prefixes, and symbols may be denoted in its SI accepted form.

[0144] Numeric ranges recited within the specification are inclusive of the numbers defining the range and include each integer within the defined range. Throughout this disclosure, various aspects of this invention are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges, fractions, and individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6, and decimals and fractions, for example, 1.2, 3.8, V >, and 43 / 4 This applies regardless of the breadth of the range.

[0145] So that the present application may be more readily understood, certain terms are first defined. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the present application pertain. Many methods and materials similar, modified, or equivalent to those described herein can be used in the practice of the embodiments of the present application without undue experimentation, the preferredmaterials and methods are described herein. In describing and claiming the embodiments of the present application, the following terminology will be used in accordance with the definitions set out below.

[0146] The term “and / or” means any one of the items, any combination of the items, or all of the items with which this term is associated.

[0147] The term “about,” as used herein, refers to variation in the numerical quantity that can occur, for example, through typical measuring techniques and equipment, with respect to any quantifiable variable, including, but not limited to, capacity, diameter, flowrate, flux, length, mass, pressure, temperature, time, and volume. Further, given typical measuring and liquid handling procedures used for making concentrates or use solutions in the real world; through inadvertent error in these procedures; through differences in the manufacture, source, or purity of the ingredients used to make the compositions or carry out the methods; and the like. The term “about” also encompasses amounts that differ due to different equilibrium conditions for a composition resulting from a particular initial mixture. Whether or not modified by the term “about,” the claims include equivalents to the quantities.

[0148] The term “capacity” as used herein refers to the amount of target component bound per unit of adsorbent.

[0149] The term “chemically stable” as used herein means that a material (including, but not limited to, a separation media) is not soluble in solvents such as water or common organic solvents (e.g., alcohols and hydrocarbons), and their mixtures.

[0150] The term “electrospinning” as used herein refers to the application of electric forces to the spin dope to form the nanofibers.

[0151] The term “exemplary” refers to an example, an instance, or an illustration, and does not indicate a most preferred embodiment unless otherwise stated.

[0152] The term “extracellular vesicles” includes vesicles of any size, including exosomes and larger extracellular vesicles. As used herein, “exosomes” refers to small, secreted vesicles (typically about 30-150 nm) which may contain, or have present in their membrane, nucleic acid, protein, or other biomolecules and may serve as carriers of this cargo between diverse locations in a body or biological system.

[0153] The term “flowrate” as used herein refers to the volume of liquid expressed in dry media volumes (MVs) flowed through membrane per time expressed in minutes. Flowrate is considered high if it is above 20 MV / min.

[0154] The term “flux” refers to the flow rate of fluid passing through the separation media per unit time, per unit of facial area exposed to the flow.

[0155] The term “fraction collector” includes any device for or means of splitting the downstream flow into fractions, whether equal in volume or dissimilar volumes. A fraction collector might comprise a stand-alone fraction collecting device equipped with collection tubes or containers or an assembly comprising valves, tubes and / or pipes to direct the downstream flow into separate storage receptacles, including but not limited to tubes, bottles, containers or tanks, or directed to waste.

[0156] The term “hybrid nanofiber felt” as used herein refers to a non-woven or randomly overlaid collection of fibers consisting of at least two types of polymers in a combination of single component fibers or composite fibers with either at least one other single component fiber or at least one other composite fiber. The term “single component nanofibers” as used herein refers to nanofibers produced from a single polymer. The term “single component nanofiber felt” as used herein refers to the accumulation of many single component nanofibers into a non-woven or randomly overlaid collection of fibers. The term “composite nanofibers” as used herein are nanofibers produced from at least two different polymers. The term “differentially removable” as used herein that, when the hybrid nanofiber felt consists of at least two non-cellulose-based polymers, conditions can be selected (elevated temperature or solvent exposure) to remove one of the non-cellulose-based polymers to a greater degree (at least 10% different, and up to 100% vs. 0%) than the other non-cellulose-based polymer.

[0157] The term “laminar” or “laminar flow” as used herein refers to flow of a liquid or gas where the liquid or gas travels smoothly or in regular paths. Laminar flow is in contrast to "turbulent flow” where the gas or liquid undergoes irregular fluctuations in its fluid path.

[0158] The term “ligand” as used herein refers to molecules or compounds capable of interaction with target compounds, such as antibodies, nucleic acids, proteins, and viruses, or particles or fragments thereof.

[0159] The term “lipid nanoparticle” or “LNP” as used herein is a general term to describe lipid-based particles in the submicron range. LNPs can have structural characteristics of liposomes and / or have alternative non-bilayer types of structures. LNPs constitute an alternative to other particulate systems, such as emulsions, liposomes, micelles, microparticles and / or polymeric nanoparticles, for the delivery of active ingredients, such as oligonucleotides and small molecule pharmaceuticals.

[0160] The term “microfibers” as used herein refers to fibers with diameters larger than 1.0 micrometer, and generally between 1.0 micrometer and 1.0 millimeter.

[0161] The term “nanofibers” as used herein refers to fibers with diameters smaller than of 1.0 micrometer, and generally between 10 nanometers and 1.0 micrometer, such as between 200 nm and 600 nm.

[0162] The term “nanofiber felt” as used herein refers to a collection of nanofibers in a substantially planar array, which may also include randomly overlaid fibers, woven fibers, non-woven fibers, electrospun nanofibrous arrays, or non-electrospun nanofibrous arrays.

[0163] As used herein, the term “nucleic acid” refers to nucleic acids in the form of deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or peptide nucleic acid (PNA), as well as analogs, derivatives, or any combination thereof. Such a derivative could contain, for example, a nucleotide analog or a “backbone” bond other than a phosphodiester bond, for example, a phosphotriester bond, a phosphoramidate bond, a phosphorothioate bond, or a thioester bond. Naturally-occurring RNA molecules include, but are not limited to, transfer RNA (tRNA), ribosomal RNA (rRNA), messenger RNA (mRNA), or genomic RNA, such as that from influenza or hepatitis C viruses. Other forms of RNA include, but are not limited to, small interfering RNA (siRNA) and microRNA (miRNA). The nucleic acid molecules can be single-stranded (ss; and which can be sense or antisense), double-stranded (ds) or a combination of the two, and can be linear or circular, the latter of which can be open-circular or closed-circular including but not limited to plasmid DNA (pDNA). The nucleic acid molecule can be a vector. The vector can be a viral vector, preferably a lentivirus vector, an adenovirus vector, an adeno-associated virus (AAV) vector, a vesicular stomatitis virus (VSV) vector, a herpes simplex virus (HSV) vector, a vaccinia virus vector, a pox virus vector, an influenza virus vector, a respiratory syncytial virus vector, a parainfluenza virus vector, a foamy virus vector, a measles virus vector or a retrovirus vector.

[0164] As used herein the term “polymer” refers to a molecular complex comprised of more than ten monomeric units and generally includes, but is not limited to, homopolymers, copolymers, such as for example, block, graft, random and alternating copolymers, terpolymers, and higher "x"mers, further including their analogs, derivatives, combinations, and blends thereof. Furthermore, unless otherwise specifically limited, the term “polymer” shall include all possible isomeric configurations of the molecule, including, but are not limited to isotactic, syndiotactic and random symmetries, and combinations thereof.Furthermore, unless otherwise specifically limited, the term “polymer” shall include all possible geometrical configurations of the molecule.

[0165] The terms “protein”, “polypeptide”, or “peptide” can be used interchangeably and refer to any natural or recombinant molecule comprising amino acids joined together by peptide bonds between adjacent amino acid residues. A “peptide bond”, “peptide link”, or “amide bond” is a covalent bond formed between two amino acids when the carboxyl group of one amino acid reacts with the amino group of the other amino acid. The terms “amino acid”, “amino acid residue”, and “residue” may be used interchangeably herein.

[0166] The terms “purifying,” “separating,” or “isolating,” as used interchangeably herein, refer to increasing the degree of purity of a molecule of interest or a target molecule from a composition or sample comprising the molecule and one or more impurities.

[0167] As used herein, the term “sample” refers to any liquid or solid material to be used in the processes described herein. For example, a sample can be a solution containing eukaryotic or prokaryotic cells or cellular material, or virus or viral material, or bacteria or bacterial material, or microorganisms or pathogens. A sample can be essentially water, or a buffered solution or be composed of any artificially introduced chemicals and may or may not contain nucleic acids or proteins.

[0168] As used herein, “biological sample” refers to any sample obtained from a living, once living, or viral source or other source of macromolecules and biomolecules, and includes any cell type or tissue of a subject from which nucleic acid or protein or other macromolecule can be obtained. The biological sample can be a sample obtained directly from a biological source or a sample that is processed. For example, isolated nucleic acids that are amplified constitute a biological sample. Biological samples can include biological solid material or biological fluid or a biological tissue. Examples of biological solid materials include tumors, cell pellets, or biopsies. Examples of biological fluids include cell cultures, cell homogenates, suspension of cells in a medium, urine, blood, plasma, serum, sweat, saliva, semen, stool, sputum, cerebral spinal fluid, mouth wash, tears, mucus, sperm, amniotic fluid, or the like. Biological tissues are aggregates of cells, usually of a particular kind, together with their intercellular substance that form one of the structural materials of a human, animal, plant, bacterial, fungal or viral structure, including connective, epithelium, muscle and nerve tissues. Examples of biological tissues also include organs, tumors, lymph nodes, arteries, and individual cell(s). Also included the definition of biological sample are soil, water andother environmental samples including industrial waste and natural bodies of water (lakes, streams, rivers, oceans) that can contain viruses, bacteria, fungi, algae, protozoa, and components thereof.

[0169] The term “spin dope” as used herein refers to the polymer solution that is used in the electrospinning process.

[0170] The term “target component” includes, but is not limited to, any particular component which is desired to be separated and captured. Preferably, “target components” are soluble components, including but not limited to, biological entities (including, but not limited to, cells, viruses, proteins, extracellular vesicles, nucleic acids, peptides, polypeptides, etc.), biocomponents, and chemical components (including, but not limited to, synthetic organics, metal ions, small molecules, etc.). In some embodiments, the target component can be found in and / or isolated from a biological sample. Preferred target components include biocomponents.

[0171] The term “thermally stable” as used herein means that a material (including, but not limited to, a separation media) does not disintegrate in the temperature range from 50-110°C.

[0172] The term “substantially” refers to a great or significant extent. “Substantially” can thus refer to a plurality, majority, and / or a supermajority of said quantifiable variables, given proper context.

[0173] A “vector” is a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Examples of vectors include but are not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “vector” includes an autonomously replicating plasmid or a virus. The term is also construed to include nonplasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, exosomes and the like. Examples of viral vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, and the like.

[0174] A “virus-like particle” or “VLP” as used herein refers to at least one virus particle, which does not contain any nucleic acid. VLPs can thus be used for vaccination or for inducing an immunogenic reaction in a subject. However, due to the absence of nucleic acids, VLPs will not be able to replicate in a host cell and are thus non-replicative.

[0175] The term “weight percent,” “wt.%,” “percent by weight,” “% by weight,” and variations thereof, as used herein, refer to the concentration of a substance as the weight of that substance divided by the total weight of the composition and multiplied by 100. It is understood that, as used here, “percent,” “%,” and the like are intended to be synonymous with “weight percent,” “wt.%,” etc.

[0176] The present disclosure is not to be limited to that described herein. Mechanical, electrical, chemical, procedural, and / or other changes can be made without departing from the spirit and scope of the present disclosure. No features shown or described are essential to permit basic operation of the present disclosure unless otherwise indicated.

[0177] Separation Device

[0178] As shown in Figure 1, a separation device 000 can comprise three main components: a lid 100, a capsule body 200, and a separation media assembly 300 integrated therewithin (the separation media 300 having a core 600 within the center void (the center void 320 shown in Figures 10A-10C); for the purpose of viewing the core 600 is upside down in reference to the separation media assembly 300 in Figure 1 (the assembled orientation of the separation media assembly 300 with the core 600 is depicted in Figures 9 and 10D). In a preferred embodiment, the separation media assembly 300 is housed at least partially within the capsule body 200.

[0179] The separation media assembly 300, shown in greater detail in Figure 8, comprises a separation media 312, a first encapsulation member 302, and optionally, but preferably, a second encapsulation member 310. The separation media 312 is pleated or alternatively rolled.

[0180] The first and second encapsulation members 302, 310, of the separation media are preferably formed via a chemical elastomeric encapsulant, such as a silicone elastomer or a polyurethane elastomer, which forms and accommodates the geometries of the separation media sealing interfaces, in addition to sealing the separation media 312. Preferably, the chemical elastomeric encapsulate has some degree of malleability and deformability. Beneficially, in preferred embodiments, the first and second encapsulation members 302, 310 seal off portions of the separation media which are encapsulated thereby reducing dead volume and forcing fluid flow through the desired portion of the separation media.

[0181] If a second encapsulation member 310 is included, it can comprise flow directing features such as channels 316 (or tapering) to provide for more efficient flow of fluid. The channels 316 are preferably grooves that the fluid can flow through thereby having a largerfluid path such that desired flow path is followed and the device can be operated at the desired pressure and speed.

[0182] The lid accommodates a first port 102; it should be understood that an aperture through the lid can form the port or house a port that travels therethrough. In some embodiments, such as those shown in Figures 2A-2B, the lid 100 also includes a ridge 104 on the underside of the lid 100. As discussed further herein, the ridge 104 aids in securing the lid 100 to the separation media assembly 300 by causing deformation of the first encapsulation member 302 which facilitates a tighter seal from fluid leakage. In a preferred embodiment, the lid can comprise a flange having an arrow-head like geometry 106 (as shown in Figures 2A and 2B) which can help secure the lid 100 to the separation media assembly 300. Interlocking features 108 of the lid 100 help secure the lid 100 to the capsule body 200. The specific geometry of the securement features, such as the ridge 104, arrow-head like geometry 106, and interlocking features 108, are non-limiting unless expressly claimed.

[0183] The capsule body includes a second port 202. In some embodiments, such as those shown in Figures 3A-3B, the capsule body 200 includes a connector 204 associated with the second port 202. The capsule body 200 can also be generally cylindrically shaped in that it can include an interior radial wall 206 and a wall 208 proximate to the second port 202. The capsule body 200 can also include a ridge 210 to help secure the capsule body 200 to the separation media assembly 300. And, in embodiments, the ridge 210 of the capsule body 200 directly opposes the ridge 104 of the lid 100 to secure a first encapsulation member 302 between the opposed ridges 210, 104. Interlocking features 216 of the capsule body 200 help secure the capsule body 200 to the lid 100. The interlocking features 216 of the capsule body 200 are designed to complement the interlocking features 108 of the lid 100 so as to facilitate securing the lid to the capsule body by engaging the interlocking features 216, 108 to each other. The specific geometry of the securement features, such as the ridge 210 and the interlocking features 216, are non-limiting unless expressly claimed. Moreover, it is to be appreciated that in some embodiments, the interlocking features 108, 216 can be permanently affixed and sonic welded to one another. In other embodiments, the capsule body and lid are separate components friction-fitted together, welded, screwed, clamped, Luer-locked, glued, adhered or otherwise secured together and sealed.

[0184] Preferably, a flow directing geometry, such as internal ridges 218, can be formed into an interior radial wall 206 of the capsule body 200 and can extend from a substantiallycylindrical portion of the capsule body 200 to an interior of the wall 208 of the capsule body 200. Alternatively, a void-volume reducing geometry can be formed into this region, such as the addition of fins, wings, tapering as further discussed below. The addition of fluid directing and void-volume reducing geometries, such as ridges, grooves, channels, tapering, fins, and wings, can aid in directing the fluid flow in intended paths and to reduce or expand the volume for fluid to travel through; this can aid in providing for equidistant paths, consistent flow times, and optimal pressure in the device.

[0185] The lid 100 has a primary interface with the separation media assembly 300, designed to hygienically couple the first port geometry with the inside region of the separation unit. Two embodiments of this interface are detailed in Figures 4-5.

[0186] A radial static seal 400 is shown in Figure 4. A connector 402, such as a Luer, a Luer- lock, a tri-clamp connection, a ferrule, and / or a threaded fitting, is associated with the first connector 402 and is directly coupled to the sealing face 404 on the underside of the lid 100.

[0187] The first encapsulation member 302 of the separation media assembly 300 preferably comprises at least one protruding portion 304 extending toward the core area of the separation unit 300. The protruding portion 304 interacts with the sealing face 404 on the underside of the lid 100, which protrudes from the lid 100 toward the core 600. The first encapsulation member 302 is preferably made of an elastomeric material and is hermetically sealed to the separation media 312. This permits a deformation of the first encapsulation member 302 so as to hermetically seal the separation media 312 and provide an intended flow path for any fluid traveling through the separation device 000. The size of connector 402 or sealing geometry is not limited unless expressly claimed.

[0188] A flange static seal is shown in Figure 5. A connector 502, such as a Luer, a Luer-lock, a tri-clamp connection, a ferrule, and / or a threaded fitting, is part of an insert that interfaces with the lid component 100. In such a design, the insert 504 compresses a sealing ring 508 that is integral to the first encapsulation member 302 between the insert flange 506 and the lid component 100. Compression is held by a retaining feature 510 and a retaining component 512 that acts against the lid 100 and may be threaded, grooved, or moulded in-place. The size of connector 502, insert flange 506, sealing geometry, retaining feature 510, or retaining component 512 are not limited unless expressly claimed. In this configuration, the connector 502 is preferably pressed through the first encapsulation 302 to compress and form the sealing ring 508 via the interaction of the flange 506.

[0189] As shown in Figures 6 and 7, the lid 100 also has a secondary interface with the separation media assembly 300 that provides the hygienic seal between the lid 100 and capsule body 200. The lid 100 preferably comprises a ridge 104 that interacts with the first encapsulation member 302. Preferably, the capsule body 200 comprises a ridge 210 that interacts with the first encapsulation member 302. The first encapsulation member 302 preferably comprises an extended portion 303 (circled in Figure 7 with 4-4), which extends beyond the diameter of the separation media 312 and is tight between the lid 100 and a lip 211 of the capsule body 200. The ridge 210 is located on the lip 211 of the capsule body 200. The ridges 104, 210 provide a pressure point on the extended portion 303 of the first encapsulation member 302 causing the first encapsulation member 302 to slightly deform and provide pressure like an O-ring (pressure inward, outward, upward and downward) thereby aiding in sealing the separation device and preventing leakage.

[0190] Figure 7 shows the integration of the separation media assembly within the capsule body 200 enclosed by the lid 100, including the first encapsulation member 302, separation media 312 and second encapsulation member 310, designed to hygienically couple the capsule body 200 with the outside region of the separation media assembly 300.

[0191] Figure 7 also shows an outer channel 212 formed by a void between the interior radial wall 206 of the capsule body 200 and the separation media 312 where the fluid can travel into along the interior radial wall 206 and into the separation media. There may be flow directing geometry, such as the ridges 218 shown in Figure 3B, or a void-volume reducing geometry (e.g., tapering) formed into this region. The interior radial wall 206 also interfaces with the second encapsulation member 310 in such a way that flow is permitted with a controlled resistance, e.g., a designed width of flow-channels, designed clearance, etc.

[0192] Figure 8 shows an embodiment of the separation media assembly 300 comprising a separation media 312 having a first encapsulation member 302 and a second encapsulation member 310. The separation media 312 is pleated in this embodiment (although it could be rolled) and join 314 is shown (in Figures 10A-10D). The second encapsulation member 310 can preferably comprise one or more fluid directing geometries (including, but not limited to, channels, ridges, grooves, and / or tapering). A preferred embodiment depicted in Figure 8 includes channels 316 where the fluid can flow around the second encapsulation member 310; these channels 316 can be in addition to ridges 218 or an alternative thereto. In a preferredembodiment, the separation device 000, 1000, 2000 comprises ridges 218, channels 316, or both.

[0193] As shown in Figure 9, the separation device 000 comprises a core 600 positioned in the center void (the center void 320 is depicted in Figures 10A-10C) of separation media 312. The core 600 can be solid or hollow, but is impermeable such that fluid does not enter the core, but flows around the core 600 according to the flow design discussed herein.

[0194] In an embodiment, the core 600 can be part of the separation media assembly 300 such that it is made from the same elastomeric material as the first and / or second encapsulation members 302, 310. In another embodiment, the core 600 is a separate component that is engaged with the separation media assembly 300 (e.g., the separation media 312 is slipped over the core 600) prior to or at the same time the separation media assembly 300 is placed within the capsule body 200. Additionally, the wall 208 interfaces with the second encapsulation member 310 in such a way that flow is permitted with a controlled resistance, e.g., a designed width of flow-channels, a designed clearance, or the like, so that the fluid is directed to the second port 202. This is further demonstrated in Figure 11A and the discussion of Figure 11A below.

[0195] Preferably, the core 600 is hygienically coupled to the lid such that the coupling is leak proof. In a preferred embodiment, the core 600 can be attached to the lid 100, or the core 600 can be separate but placed on the lid 100 and pressed into the separation media 312 which can aid in securing and sealing the separation media 312 within the capsule body 200 with the lid 100 attached. This is particularly true of the sealing configuration depicted in Figures 4 and 6. While not depicted in the present figures, the core 600 can also be a non- cylindrical shape, such as a star or multi-pointed shape corresponding to the pleating of the separation media 312; in such an embodiment, there would still be space maintained between the core 600 and the separation media 312 such that an inner channel 214 is maintained therebetween.

[0196] The core 600 provides multiple benefits, including, but not limited to, reducing dead volume and providing an equidistant flow-path for fluid to travel around and into the separation media. The core 600 can include a radial flow region or diverter 602, flow channels, tapering, and / or other geometries to aid fluid flow or reduce dead volume. For example, in some embodiments, particularly, larger volume designs, the core 600 cancomprise fins and / or vents, which can provide additional structural support for the separation media, reduce dead volume, and / or aid the fluid flow path.

[0197] As shown in Figures 10A-10D, the separation media 312 is preferably pleated (although it could be rolled) and forms a cylinder or other shape that matches the shape of the capsule body 200. While Figures 10A-10D utilize reference characters 312, 3214, 318A, 318B, 320, it should be understood that it is also applicable to the separation devices 1000 and 2000 and not limited to separation device 000. Thus, the corresponding numbers 312 depicted in Figures 10A-10D should be understood to include 1312 and 2312 and so forth. The number of pleats can be influenced by the size of the capsule body 200 and the thickness of the separation media 312. Preferably, there are at least 6 pleats, at least 7 pleats, at least 8 pleats, at least 9 pleats, at least 10 pleats, at least 11 pleats, at least 12 pleats, at least 13 pleats, at least 14 pleats, at least 15 pleats, at least 16 pleats, at least 17 pleats, at least 18 pleats, at least 19 pleats, at least 20 pleats, at least 21 pleats, at least 22 pleats, at least 23 pleats, at least 24 pleats, at least 25 pleats, at least 26 pleats, at least 27 pleats, at least 28 pleats, at least 29 pleats, at least 30 pleats. Most preferably there are at least 12 pleats or at least 20 pleats.

[0198] Figures 10A-10C show that the separation media 312 (and correspondingly 1312 and 2312) comprises a center void 320, Figure 10C. Figure 10D shows that the core 600 is positioned in the center void 320.

[0199] Figures 10A-10D also demonstrates that the pleated separation media 312 may comprise a join 314. The join is bonded (e.g., chemically and / or thermally). The join can also be encapsulated as shown in Figure 10C; in such an embodiment, the encapsulation can serve as a chemical bond. In some embodiments, it is possible for the separation media 312 to be manufactured as cylindrical membrane (and subsequently folded into pleats) or a cylindrical, pleated membrane; for example, via electrospinning or casting. More preferably, the separation media is manufactured in one or more sheets, which are then formed into the desired pleated configuration and the ends of the one or more sheets are formed together to form a join, which is thermally bonded, chemically bonded, and / or encapsulated. Where an encapsulated join 314 is utilized, the encapsulation is preferably formed of a chemically similar compound which hermitically seals with the first and second encapsulation members 302, 310. The separation media 312 can comprise multiple joins, where more than one sheet are utilized to form the separation media 312; thus, the separation media 312 can comprise 1j oin, 2 j oins, 3 j oins, 4 j oins, 5 j oins, 6 j oins, 7 j oins, 8 j oins, 9 j oins, 10 j oins or more than 10 joins. Most preferably, the separation media 312 comprises 1, 2, or 3 joins.

[0200] Figure 10B shows that the separation media 312 preferably comprises a primary functional layer 318A which comprises the surface functionalized separation media which may be a nanofiber material and optionally a secondary inert layer 318B. The optional secondary inert layer 318B can be added to aid in separating the separation media from itself along the pleats and can also provide rigidity and physical support (including, but not limited to, limiting or preventing deformation or warping of the separation media under high pressure conditions); the secondary inert layer 318B is not biologically or chemically reactive such that it does not participate in the capture chemistry. In a preferred embodiment, the secondary inert layer 318B is a spunbond nonwoven layer, including, but not limited to, a non-woven polypropylene. Alternatively, the secondary inert layer may be a woven polymer fabric or mesh. Additionally, layer types may be added or removed for different embodiments either external to the secondary inert layer 318B or between the secondary nonfunctional inert layer 318B and the primary functional layer 318A of the separation media 312. In an embodiment, a secondary nonfunctional inert layer is between two primary functional layers on either side such that it is ‘sandwiched’ between the primary functional layers. Accordingly, the separation media can comprise at least two layers, at least three layers, at least four layers or more. In such an embodiment, more than one join may be utilized as appropriate.Separation Media

[0201] The separation media comprises a primary functional layer 318A, which can be any suitable media for performing a separation. Accordingly, the separation devices described herein are not limited by the type of separation media or separation / capture technique. Preferred separation media include hydrogels, membranes, monoliths, matrices, or other porous structures. In a preferred embodiment, the separation media comprises a cast membrane. In another preferred embodiment, the separation media comprises a fibrous membrane. Fibrous membranes can comprise one or more fibers woven or randomly overlaid such that channels and / or pores are formed. The one or more fibers can be microfibers, nanofibers, of a diameter larger than microfibers, or a combination thereof. Preferably, the separation media comprises channels and pores. The channels and / or pores can be substantially uniform in diameter or be of varying diameters. Most preferably, the separation media is suitable for use inbioseparations. Regardless of whether the separation media 312, 1312, 2312 is rolled or pleated, it can comprise one or more layers of membrane and / or inert material.

[0202] If utilizing a membrane, preferred membranes include, but are not limited to, electrospun membranes, cast membranes, fiber felts, nanofiber felts and hybrid felts. In a most preferred embodiment, a membrane can comprise hybrid felts composed of electrospun nanofibers. Preferred membranes are described in U.S. Pat. No. 9,604,168 and PCT / US17 / 30078, the detailed disclosures of each are incorporated herein by reference in their entirety. Hybrid nanofiber felts have a high separation capacity and provide reproducible performance over multiple cycles under both high flow and high pressure. Such nanofiber felts exhibit complex interconnected, three-dimensional porous structures and relatively large surface areas.

[0203] Preferred felts comprise more than one polymer type ( / .< ., they are “hybrid” felts.) This includes hybrid felts made from a combination of single component nanofibers and “composite” nanofibers (e.g., the nanofibers are made from a mixture of two or more materials) into the “hybrid” felt. For the “composite” nanofiber, the “backbone polymer” is a derivatized cellulose, and the first non-cellulosic polymer is capable of being removed from the fiber / felt by exposing it to an elevated temperature or chemical solvents, or both an elevated temperature and chemical solvents. In some embodiments, the removal of the first non-cellulosic polymer simultaneously converts the derivatized cellulose back to cellulose, i.e., the cellulose is “regenerated.” In some embodiments the first non-cellulosic polymer is not capable of being removed and composite nanofibers incorporated into felts may comprise a backbone polymer and one or more non-cellulosic polymers.

[0204] The nanofibers in these felts are preferably manufactured using an electrospinning technique. This refers to the manufacture of fibers based on exposure of an extruded polymer “spin dope” to an electrostatic field which results in elongation of the extruded polymer “jet” into a nanofiber.

[0205] In a preferred embodiment, the separation media comprises a fibrous membrane and is particularly suitable for use in bioseparations and exhibits one or more of the following technical features: (1) small diameter fibers to allow for the largest amount of specific area; (2) well-controlled and narrow pore size distribution between fibers to allow for even flow distribution during adsorptive applications; (3) fibers having excellent mechanical and chemical stability to withstand potentially high operating pressures and harsh cleaningconditions; and (4) fibers having a well-defined and spatially consistent size and chemical composition. In a more preferred embodiment, the membrane support comprises crosslinked fibers. In a most preferred embodiment, the membrane support comprises crosslinked cellulose nanofibers; such as the crosslinked nanofibers described in U.S. Patent No. 10,919,986, which is incorporated by reference in its entirety.

[0206] The separation media can be comprised of any suitable material; preferably, it is a material that can be surface functionalized as described further below. Preferred materials include, but are not limited to, natural polymers and synthetic polymers. Preferred natural polymers include, but are not limited to, collagen, chitosan, agarose, agarose acetate, cellulose, cellulose acetate, crosslinked cellulose, derivatized cellulose, regenerated cellulose, and combinations thereof. Preferred synthetic polymers include, but are not limited to, those comprised of vinyl polymers, acrylic polymers, and copolymers thereof. More preferred synthetic polymers include, but are not limited to, polyacrylic acid, polymethacrylic acid, polyacrylonitrile (PAN), polyethylene oxides, polyimides, polyamides (nylon 6, nylon 6,6, nylon 6,10, etc.), polyesters (polyethylene terephthalate, etc.), polystyrene, polysulfones, polycaprolactone, and copolymers thereof.

[0207] Preferred polymers can include, but are not limited to, thermoplastic homopolymers such as vinyl polymers, acrylic polymers, polyamides, polyesters, polyethers, and polycarbonates including, but not limited to, polyacrylic acid, polymethacrylic acid, nylon, polyethersulfone, and polyacrylonitrile, (2) thermoplastic copolymers such as vinyl-co-vinyl polymers, acrylic-co-acrylic copolymers and vinyl-coacrylic polymers, (3) elastomeric polymers such as triblock copolymer elastomers, polyurethane elastomers, and ethylene- propylene-diene-elastomers, (4) high performance polymers such as polyimides and aromatic polyamides, (5) liquid crystalline polymers such as poly(p-phenylene terephthalamide) and polyaramid, (6) textile polymers such as polyethylene terephthalate and polyacrylonitrile, (7) electrically conductive polymers such as polyaniline, as well as (8) biocompatible polymers (i.e. “biopolymers”) like poly caprolactone, polylactide, chitosan and polyglycolide. As described, the polymer may also be a copolymer of two or more of the above-named polymer species.

[0208] Surface Functionalization

[0209] For use in bioseparation, the separation media is ideally biologically inert, meaning it generally resists non-specific binding of insoluble solids such as cells and cellular debris, aswell as unwanted interactions with proteins, sugars, nucleic acids, viruses, and other soluble components present in many biologically produced systems.

[0210] Preferably, the separation media comprises one or more surface functionalizations that aid in separation of one or more target components. The surface functionalizations can be part of the separation media or attached to the separation media. Any suitable surface functionality for a particular separation technique can be utilized with a compatible separation media. Preferred surface functionalities include, but are not limited to, ion exchange groups, hydrophobic groups, chelating groups, affinity ligands, mixed mode ligands (also referred to as multi-modal ligands).

[0211] Target “biocomponents” include, but are not limited to, biomolecules and fragments thereof. Preferred target biocomponents, include, but are not limited to, an amino acid, a peptide, an affimer, a biomolecule and / or fragment thereof, a protein, an enzyme, a glycoprotein, a lipopolysaccharide, an antibody and / or a fragment thereof, an LNP, a nucleic acid, an organic polymer, a virus, a VLP, an extracellular vesicle (EV), an exosome, a bacterium, a cell, and a cell-related structure. Preferred antibodies include but are not limited to, monoclonal antibodies, single-chain antibodies, bi-specific antibodies, multi-specific antibodies, antibody conjugates, antibody fusion proteins, drug-antibody conjugates, antibody fragments, Fab fragments, Fv fragments, gamma globulins (IgG), IVIG, IgM, IgA, IgE, hyperimmune gamma globulins and isoagglutinins. Preferred viruses, include, but are not limited to, adeno-associated virus (AAV), measles virus and lentivirus. Preferred cells and cell- related structures are derived from plants or animals; most preferably cells and cell-related structures are animal derived.

[0212] The surface functionalizations can be present at density of from about 0.001 mg / g to about 999 mg / g. Embodiments provide the ligand can be present at density of from about 0.001 mg / g to about 500 mg / g, from about 0.01 mg / g to about 400 mg / g, from about 0.1 mg / g to about 300 mg / g, from about 0.5 mg / g to about 200 mg / g, from about 1 mg / g to about 100 mg / g, about 10 mg / g to about 100 mg / g, from about 10 mg / g to about 200 mg / g, from about 10 mg / g to about 250 mg / g, from about 10 mg / g to about 300 mg / g, from about 10 mg / g to about 400 mg / g, from about 10 mg / g to about 500 mg / g, from about 10 mg / g to about 600 mg / g, from about 10 mg / g to about 700 mg / g, from about 10 mg / g to about 750 mg / g, from about 10 mg / g to about 800 mg / g, from about 10 mg / g to about 900 mg / g, from about 50 mg / g to about 100 mg / g, from about 50 mg / g to about 200 mg / g, from about 50 mg / g to about250 mg / g, from about 50 mg / g to about 300 mg / g, from about 50 mg / g to about 400 mg / g, from about 50 mg / g to about 500 mg / g, from about 50 mg / g to about 600 mg / g, from about 50 mg / g to about 700 mg / g, from about 50 mg / g to about 750 mg / g, from about 50 mg / g to about 800 mg / g, from about 50 mg / g to about 900 mg / g, from about 50 mg / g to about 999 mg / g, from about 100 mg / g to about 200 mg / g, from about 100 mg / g to about 250 mg / g, from about 100 mg / g to about 300 mg / g, from about 100 mg / g to about 400 mg / g, from about 100 mg / g to about 500 mg / g, from about 100 mg / g to about 600 mg / g, from about 100 mg / g to about 700 mg / g, from about 100 mg / g to about 750 mg / g, from about 100 mg / g to about 800 mg / g, from about 100 mg / g to about 900 mg / g, from about 100 mg / g to about 999 mg / g, from about 200 mg / g to about 250 mg / g, from about 200 mg / g to about 300 mg / g, from about 200 mg / g to about 400 mg / g, from about 200 mg / g to about 500 mg / g, from about 200 mg / g to about 600 mg / g, from about 200 mg / g to about 700 mg / g, from about 200 mg / g to about 750 mg / g, from about 200 mg / g to about 800 mg / g, from about 200 mg / g to about 900 mg / g, from about 200 mg / g to about 999 mg / g, from about 250 mg / g to about 300 mg / g, from about 250 mg / g to about 400 mg / g, from about 250 mg / g to about 500 mg / g, from about 250 mg / g to about 600 mg / g, from about 250 mg / g to about 700 mg / g, from about 250 mg / g to about 750 mg / g, from about 250 mg / g to about 800 mg / g, from about 250 mg / g to about 900 mg / g, from about 250 mg / g to about 999 mg / g, from about 300 mg / g to about 400 mg / g, from about 300 mg / g to about 500 mg / g, from about 300 mg / g to about 600 mg / g, from about 300 mg / g to about 700 mg / g, from about 300 mg / g to about 750 mg / g, from about 300 mg / g to about 800 mg / g, from about 300 mg / g to about 900 mg / g, from about 300 mg / g to about 999 mg / g, from about 400 mg / g to about 500 mg / g, from about 400 mg / g to about 600 mg / g, from about 400 mg / g to about 700 mg / g, from about 400 mg / g to about 750 mg / g, from about 400 mg / g to about 800 mg / g, from about 400 mg / g to about 900 mg / g, from about 400 mg / g to about 999 mg / g, from about 500 mg / g to about 600 mg / g, from about 500 mg / g to about 700 mg / g, from about 500 mg / g to about 750 mg / g, from about 500 mg / g to about 800 mg / g, from about 500 mg / g to about 900 mg / g, from about 500 mg / g to about 999 mg / g, from about 600 mg / g to about 700 mg / g, from about 600 mg / g to about 750 mg / g, from about 600 mg / g to about 800 mg / g, from about 600 mg / g to about 900 mg / g, from about 600 mg / g to about 999 mg / g, from about 700 mg / g to about 750 mg / g, from about 700 mg / g to about 800 mg / g, from about 700 mg / g to about 900 mg / g, from about 700 mg / g to about 999 mg / g, from about 750 mg / g to about 800 mg / g, from about 750 mg / g to about 900 mg / g, from about 750 mg / g to about 999mg / g, from about 800 mg / g to about 900 mg / g, from about 800 mg / g to about 999 mg / g, or from about 900 mg / g to about 999 mg / g. Embodiments provide the ligand may be present at density of from about 10 mg / g, about 50 mg / g, about 100 mg / g, about 200 mg / g, about 250 mg / g, about 300 mg / g, about 400 mg / g, about 500 mg / g, about 600 mg / g, about 700 mg / g, about 750 mg / g, about 800 mg / g, about 900 mg / g, or about 999 mg / g.

[0213] Preferably, the surface functionalized separation media has a dynamic binding capacity on a volume basis between about 1 pg / mL and about 400 mg / ml of the separation media; more preferably between about 5 pg / mL and about 400 mg / mL of the separation media. It should be understood that the binding capacity can vary depending the target molecule. For example, if the targe molecule is a small molecule or microscopic material (e.g., virus) then the amount captured compared to the mass of separation media will be small. However, in some applications the binding capacity can be greater due to capture of a larger molecule. Accordingly, in a some embodiments, the separation media preferably has a binding capacity on a volume basis of at least 1 pg / mL of the separation media, at least about 5 pg / mL of the separation media, at least about 10 pg / mL of the separation media, at least about 15 pg / mL of the separation media, at least about 20 pg / mL of the separation media, at least about 25 pg / mL of the separation media, at least about 30 pg / mL of the separation media, at least about 35 pg / mL of the separation media, at least about 40 pg / mL of the separation media, at least about 45 pg / mL of the separation media, at least about 50 pg / mL of the separation media, at least about 55 pg / mL of the separation media, at least about 60 pg / mL of the separation media, at least about 65 pg / mL of the separation media, at least about 70 pg / mL of the separation media, at least about 75 pg / mL of the separation media, at least about 80 pg / mL of the separation media, at least about 85 pg / mL of the separation media, at least about 90 pg / mL of the separation media, at least about 95 pg / mL of the separation media, at least about, 100 pg / mL of the separation media, at least about 110 pg / mL of the separation media, at least about 120 pg / mL of the separation media, at least about 130 pg / mL of the separation media, at least about 135 pg / mL of the separation media, at least about 140 pg / mL of the separation media, at least about 150 pg / mL of the separation media, at least about 160 pg / mL of the separation media, at least about 170 pg / mL of the separation media, at least about 180 pg / mL of the separation media, at least about 190 pg / mL of the separation media, at least about 200 pg / mL of the separation media, at least about 250 pg / mL of the separation media, at least about 300 pg / mL of the separation media, at leastabout 350 pg / mL of the separation media, at least about 400 pg / mL of the separation media, at least about 450 pg / mL of the separation media, at least about 500 pg / mL of the separation media, at least about 550 pg / mL of the separation media, at least about 600 pg / mL of the separation media, at least about 650 pg / mL of the separation media, at least about 700 pg / mL of the separation media, at least about 750 pg / mL of the separation media, at least about 800 pg / mL of the separation media, at least about 850 pg / mL of the separation media, at least about 900 pg / mL of the separation media, at least about 950 pg / mL of the separation media, at least about 1 mg / mL of the separation media, at least about 2 mg / mL of the separation media, at least about 3 mg / mL of the separation media, at least about 4 mg / mL of the separation media, at least about 5 mg / mL of the separation media, at least about 6 mg / mL of the separation media, at least about 7 mg / mL of the separation media, at least about 8 mg / mL of the separation media, at least about 9 mg / mL of the separation media, at least about 10 mg / mL of the separation media, at least about 15 mg / mL of the separation media, at least about 20 mg / mL of the separation media, at least about 25 mg / mL of the separation media, at least about 30 mg / mL of the separation media, at least about 35 mg / mL of the separation media, at least about 40 mg / mL of the separation media, at least about 45 mg / mL of the separation media, at least about 50 mg / ml of the separation media, at least about 55 mg / ml of the separation media, at least about 65 mg / ml of the separation media, at least about 70 mg / ml of the separation media, at least about 75 mg / ml of the separation media, at least about 80 mg / ml of the separation media, at least about 85 mg / ml of the separation media, at least about 90 mg / ml of the separation media, at least about 95 mg / ml of the separation media, at least about 100 mg / ml of the separation media, at least about 110 mg / ml of the separation media, at least about 120 mg / ml of the separation media, at least about 130 mg / ml of the separation media, at least about 140 mg / ml of the separation media at least about 150 mg / ml of the separation media.

[0214] Preferably, the surface functionalized separation media has a dynamic binding capacity for proteinaceous target components on a mass basis of at least about 10 mg / g of the separation media in hydrated form, at least about 95 mg / g of the separation media, at least about 100 mg / g of the separation media, at least about 105 mg / g of the separation media, at least about 110 mg / g of the separation media, at least about 115 mg / g of the separation media, at least about 120 mg / g of the separation media, at least about 125 mg / g of the separation media, at least about 130 mg / g of the separation media, at least about 125 mg / g ofthe separation media, at least about 130 mg / g of the separation media, at least about 135 mg / g of the separation media, at least about 140 mg / g of the separation media, at least about 145 mg / g of the separation media, at least about 150 mg / g of the separation media, at least about 155 mg / g of the separation media, at least about 160 mg / g of the separation media, at least about 165 mg / g of the separation media, at least about 170 mg / g of the separation media, and at least about 175 mg / g of the separation media.

[0215] Preferably, the surface functionalized separation media has a dynamic binding capacity for nucleic acid components on a mass basis of at least about 2 mg / mL of separation media, at least about 5mg / mL of separation media, at least about lOmg / mL of separation media, at least about 15mg / mL of separation media and at least about 20mg / mL of separation media.

[0216] In the case of virus particles or VLPs, binding capacity can be measured based on the number of particles bound per mL of separation media. Accordingly, binding capacity for virus particles and VLPs is between about 1 x 1010particles per mL and about 1 x 1011particles per mL, more preferably between about 1 x 1010particles per mL and about 1 x 1012particles per mL, still more preferably between about 1 x 1010particles per mL and about 1 x 1013particles per mL, between about 1 x 1010particles per mL and about 1 x 1014particles per ml and most preferably between about 1 x 1012particles per mL and about 1 x 1014particles per mL.

[0217] Ion Exchange Surface Functionalizations

[0218] Suitable ion exchange surface functionalizations include, but are not limited to, anion exchange groups, cation exchange groups, and mixtures thereof.

[0219] Anion exchange groups contain one or more moieties which are positively charged and attract anionic molecules. Preferred anion exchange groups include, but are not limited to, tertiary and quaternary amines. Preferred anion exchange groups include, but are not limited to, amine based moieties, including but not limited to quaternary ammonium groups such as alkyltrimethylammonium groups (sometimes referred to as Q groups), diethyl-(2-hydroxy- propyl), aminoethyl groups (sometimes referred to as a QAE exchange group), trimethylaminoethyl groups (sometimes referred to as TMAE groups), tertiary amine groups such as diethylaminoethyl (DEAE), and mixtures thereof. Preferred amine based moieties include, but are not limited to — N+(CH3)3, — N+(C2H5)H, — CEECEEN^CEE — O —CH2CH2— N+(CH3)3, — CH2CH2N+(C2H5)H, — CH2CH2N+(C2H5)2(CH2CH(OH)CH3), and — CH2CH2N+(CH3)2H moieties.

[0220] Cation exchange groups contain one or more moieties which are negatively charged and attract cationic molecules. Preferred cation exchange groups include, but are not limited to, one or more carboxylate ( — COO-), sulphonate ( — SO3-), phosphonate ( — P(OH)2O-) groups, or mixtures thereof. Preferred weak cation exchange groups include one or more of — CH2COO-, — O-CH2COO-, — CH2OCH2COO- (sometimes referred to as a CM ion-exchange group). Preferred strong cation exchange groups include one or more of — SO3- (sometimes referred to as an S ion-exchange group), — CH2CH2CH2SO3-, (sometimes referred to as an SP ion-exchange group) — CH2CH2SO3-, — P(OH)2O-, and mixtures thereof.

[0221] Hydrophobic Surface Functionalizations

[0222] Hydrophobic surface functionalizations are used for separating molecules based on their relative hydrophobicity. Surface functional groups suitable for this include one or more hydrophobic moieties. Preferred hydrophobic moieties include, but are not limited to, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, octyl, decyl or phenyl groups, or mixtures thereof.

[0223] Affinity Ligand Surface Functionalizations

[0224] Affinity ligand surface functionalizations separate molecules based on their affinity to particular ligands. In a preferred embodiment, an affinity ligand is a chemical or biological ligand, which for example, can rely on a multiplicity of attractive forces including electrostatic, hydrophobic, hydrogen bonding and Van der Waal’s forces in combination with an advantageous spatial orientation of binding groups as exists, for example, in the case of the binding of antibodies to antigens, protein A to IgG or substrates and inhibitors to enzymes.

[0225] Suitable affinity ligands include, but are not limited to, chemical ligands, dyes, small molecule biomimetic ligands, triazine ligands, immobilized metal ions, affimers, aptamers, affibodies, peptides, polypeptides, antibodies, enzyme inhibitors, lectins and bacterial immunoglobulin binding proteins.

[0226] Preferred affinity ligands include, but are not limited to, monoclonal antibodies, polyclonal antibodies, antibody fragments, dyes, histidine, a group comprising a metal ion, nucleic acids, oligonucleotides, lectins, proteins, oligopeptides, polysaccharides, oligosaccharides, sugars, peptides, antigens, aptamers, affimers, small organic compounds, synthetic affinity ligands (typically with molecular weights <2,000 including, but not limitedto, triazine ligands, aminophenyl boronate, and aminobenzamidine), drugs and other ligands. Examples of suitable affinity ligands are available in the published literature and are well known.

[0227] In a preferred embodiment the ligand can bind an immunoglobulin. Non-limiting, examples of such ligands include Protein A, Protein G, and Protein L. As used herein, “Protein A” refers to recombinant Protein A (which may have an altered sequence compared to Protein A found in Staphylococcus aureus) and tagged Protein A (as described in EP-B- 0873353 and U.S. Pat. No. 6,399,750, the detailed disclosures of which are incorporated herein by reference). Protein A may be a modified variant of Protein A, for instance cysteine modified variants of Protein A.

[0228] Small molecule biomimetic ligands are small molecule chemical ligands with a molecular mass < 2,000 D that mimic the binding of interactions of naturally occurring ligands.

[0229] A preferred antibody and antibody fragment includes, but is not limited to, IgG, IgM, monoclonal antibodies, camelid antibodies, antibody fragments, Fab, Fv, and combinations thereof.

[0230] Certain dyes can be employed as affinity ligands. Preferred dyes include, but are not limited to, Procion Blue HB, Procion Yellow HE-4R, Procion Red HE-3B, and Cibacron Blue F3G.

[0231] Another category of compounds useful as affinity ligands includes groups containing a metal ion, i.e., an immobilized metal ion. Preferred groups containing metal cations are chelating groups which contain and immobilize one or more metal cations. Preferably the metal cation contained is one or more of copper, nickel, zinc, iron and cobalt; most preferably Cu2+, Ni2+, Zn2+, Fe3+, Co2+or a combination thereof.

[0232] Mixed Mode Ligand Surface Functionalizations

[0233] Mixed mode ligands (sometimes referred to as multimodal ligands) utilize two or more types of binding interaction via a single ligand, i.e., a single ligand possessing at least two of an anion exchange group, a cation exchange group, a polar group, a hydrophobic group or a hydrogen bonding group, which separates molecules based on two or more characteristics of the target molecule. In a preferred embodiment, a mixed mode ligand comprises an ion exchange group in addition to a hydrophobic group or a hydrogen bonding group. In a preferred embodiment, a mixed mode ligand comprises a hydrophobic group inaddition to a hydrogen bonding group, a polar group, or an ion exchange group. In a preferred embodiment, a mixed mode ligand comprises a hydrogen bonding group in addition to an ion exchange group or hydrophobic group. Preferred mixed mode ligands include, but are not limited to, those comprising an N-benzyl methyl ethanolamine group, an N-benzoyl- homocysteine group, or combinations thereof.

[0234] A surface functionalized separation media for use in such methods may also contain one or more hydrophobic groups which are ionizable, for use in so-called Hydrophobic Charge Induction Chromatography (HCIC). Thus, in one embodiment, a mixed mode ligand comprises a HCIC compatible group. Preferred mixed mode ligands suitable for suitable for HCIC include, but are not limited to, 4-mercapto-ethyl-pyridine (MEP) groups and octylamine groups.

[0235] Methods of Using the Separation Device

[0236] As shown in Figure 11 A, a fluid enters the separation device 000 via the first port 102 and fluid is directed by a radial flow region 602 (or optionally a diverter) and the underside of the first encapsulation member 302 to travel around the core 600 to the inner channel 214 which is adjacent to an interior surface of the separation media 312. The inner channel 214 is a void space between the wall of the core 600 and the separation media 312. Fluid passes through the separation media 312 radially toward an outer channel 212 formed by the void between the exterior surface of the separation media 312 and the interior radial wall 206 of the capsule body 200. The fluid then travels through the outer channel 212 of the capsule body 200 toward the second port 202. In some embodiments, a second encapsulation member 310 is included, which can comprise channels 316 permitting fluid to flow with less resistance around the second encapsulation member 310 and to the second port 202.

[0237] In a preferred embodiment, the outer channel 212 tapers from a first end of the outer channel 212A which has a narrower width and is near the lid 100 and first encapsulation member 302 to a second end of the outer channel 212B which has a wider width and is near the second encapsulation member 310. Conversely, an inner channel 214 tapers from a wider width at a first end 214A near the lid 100 to a narrower 214B near the end proximate the second port 202. The converse tapering of the outer channel 212 and inner channel 214 is preferably at a corresponding slope such that this helps maintain the geometry provisioning equidistant paths through the unit, and therefore, the equivalent residence-time within thedevice where laminar flow is concerned. By providing this, the flow can be time matched between the flow of liquid in the inner channel 214 and the outer channel 212.

[0238] The narrowing inner channel 214 is designed to improve the fluid flow characteristics (i.e., low pressure, high flow rate, and low residence time) so that there is more space to accommodate the fluid coming from the first port; as fluid enters the separation media 312 less of a volume is needed. Moreover, by narrowing the inner channel 214 in the direction of the fluid path, fluid that has not yet entered the separation media can be forced into the separation media.

[0239] The widening outer channel is designed to provide the same fluid flow characteristics (i.e., low pressure, high flow rate, and low residence time). However, because this is the channel the fluid will exit from, there will be less fluid at the first end 212A. As fluid disperses radially through the separation media 312 into the outer channel 212 the volume will increase as it travels toward the second port; thus, by increasing the width of the outer channel along the flow path, the volume of fluid that can travel toward the second port can be increased. These inverse tapering features of the inner channel and outer channels can provide for a low pressure, while maintaining high flow rates and low residence times.

[0240] Figure 11B is an enlarged view of the lower right corner portion of Figure HA such that the flow channels 604 on the second encapsulation member 310. As can be seen in Figure 11B, fluid flow is permitted to flow around the second encapsulation member 310 in the outer channel 212 and then under the second encapsulation member 310 through one or more flow channels 604.

[0241] Most preferably, the relationship between the tapering is such that equidistant paths are provided throughout the separation device 000. This is in part represented and demonstrated in Figure 11C, where laminar flow is observed via example flow-lines, each has an equivalent residence-time within the device. This results from the specific geometry of the device providing equidistant paths through the unit. Where turbulent flow may exist, this is not limited to always being the case as the fluid no longer travels in an equidistant path but in an irregular path. It should be understood that depending on the separation device configuration, the lateral flow arrows can extend externally (representing flow through the separation media radially outwardly) as depicted in Figure 11C or can extend in a reverse direction, i.e., the radial flow traveling inwardly through the separation media; under either flow path, the laminar flow and equidistance of travel are achieved.

[0242] In another embodiment, the flow can be reversed such that the fluid enters from the second port 202 near the first encapsulation member 302 and travels toward the interior radial wall 206 and passing through the outer channel 212 where the fluid enters the separation media and travels inward toward the inner channel 214 until it reaches the tapering inner flow path boundary where it flows toward the second end of the inner channel 214B and the second port 202. While some of the benefits of the membrane could still be utilized in such a process, many of the benefits related to flow speed, pressure, and residence time would not be achieved in this reverse flow. Rather, this reverse flow may be primarily used to aid in removing any particulate matter from the inner diameter of the separation media which can accumulate there depending on the nature of the fluids traveling through the separation device.

[0243] Figure 11D shows a detailed perspective view of a preferred separation media assembly 300 having a second encapsulation member 310, a separation media 312, and a first encapsulation member 302. In this embodiment, the second encapsulation member 310 comprises optional flow channels 604, which provide a path for fluid to flow after traveling through the separation media 312 and around the second encapsulation member 310 to reach the second port 202. In addition to or as an alternative to the flow channels 604, ridges 218 can be formed in the capsule body 200 interior to the wall 208 to provide a path for the fluid to travel; thus, in a preferred embodiment, the separation device 000, 1000, 2000 comprises ridges 218, flow channels 604, or both. Using the radial spoke design depicted in Figure 11D, whether via channels 604 and / or ridges 218 (Figure 11D shows channels, but the same design can be used in the interior capsule body as ridges 218), can aid in providing laminar flow and thus more uniform fluid flow toward the second port 202. The flow channels 604 can be in a variety of configurations to achieve different fluid flows and are not limited to the radial spoke style design depicted in Figure 11D. Moreover, other flow directing geometries can be provided to direct the fluid flow toward the second port 202 including, but not limited to, tapering, use of legs or other geometries to provide a gap between the second encapsulation and the interior capsule body. Also visible in Figure 11D is a join 314, which has been bonded, and the second encapsulation member 310 having optional channels 316.

[0244] The channels 316 (depicted in Figures 8 and 11D) can be configured to correspond with the internal ridges 218 of the capsule body 200 (depicted in Figures 3A and 3B). Thechannels 316 can be used to provide preferential flow paths for the fluid, while the second encapsulation member 310 is tight to the interior radial wall 206.

[0245] As shown in Figure HE, a fluid enters the embodiment separation device 000 through the second connector 204 via the second port 202 and fluid diverges upon coming in contact with the second encapsulation member 310 to travel around the second encapsulation member 310. As shown in Figure HE, the second encapsulation member 310 is the same as that shown in Figure HA in which ridges 604 guide a fluid path of the fluid. However, different from Figure HA, fluid enters the separation device 000 at the second connector 204 through the second port 202 such that the separation media assembly 300 is in a reverse orientation. As such, the fluid path flows opposite to that shown in Figure HA, and the fluid coming in contact with the ridges 604 and / or channels 218 after entering the second port 202 travel radially outward rather than radially inward. The second encapsulation member 310 need not comprise ridges 604 to guide the fluid but instead could be shaped as a dome. The dome could range from that of a shallow dome (barely noticeable that the second encapsulation member 310 is not flat) to that of a half-circle, wherein the fluid may spread evenly over a surface of the dome due to its symmetry.

[0246] After traveling over the second encapsulation member 310 the fluid may then travel to the outer channel 212 formed by the void between the exterior surface of the separation media 312 and the interior radial wall 206 of the capsule body 200. The fluid may begin at the second end of the outer channel 212B and travel towards the first end of the outer channel 212A while simultaneously traveling radially inward through separation media 312. Fluid passes through the separation media 312 and into the inner channel 214 which is adjacent to an interior surface of the separation media 312 and the core 600. The fluid may then travel down the inner channel 214 around the core 600 to the underside of the first encapsulation member 302. As fluid travels between an end of the core 600 and the lid 100, the fluid will travel down and out of the first port 102.

[0247] Further included as an optional feature is a bleed-valve 450 adjacent the second port 202. The bleed-valve 450 includes a screw 452 and a bleed-valve aperture 222 as shown in Figure HE. The bleed-valve 450 allows for releasing of any air that may get caught within the separation device 000. This is advantageous as a user may find small amounts of air enter the device 000 when connecting it to a system with flows lines (infra 702, 704) that can be part of a chromatography work station, an FPLC, an Akta system, a process chromatographyskid, a membrane chromatography skid, etc. The bleed-valve 450 makes such transitions possible while maintaining the ability to have smooth fluid flow and removal of any blockages due to trespassing air. In preferred embodiments of the separation device 000, 1000, 2000 the optional bleed-valve 450 is included.

[0248] Figure 11F shows a fluid entering the embodiment of the separation device 000 via the second port 202 and fluid diverges upon coming in contact with the second encapsulation member 310 to travel around the second encapsulation member 310. However, unlike in Figure HE, the separation device 000 includes a first frit 800 and a second frit 850; as noted above, the first and second frits 800, 850 can be utilized in any of the separation devices 000, 1000, 2000 described and shown herein. Fluid still enters the separation device 000 at the second port 202, but the fluid travels through the first frit 800 before traveling around the second encapsulation member 310. The first frit 800 can act as a filter, provide direction and dispersion of fluid, and / or used to reduce dead-volume. Should any foreign material or particles exist within the fluid feed, the frit’s porous structure can assist in preventing fine particles from traveling through the separation device 000. Additionally, the first frit 800 can assist in maintaining the stationary phase. The first frit 800 can also assist in uniform distribution of the mobile phase. Frits can be made of sintered polyethylene, sintered polypropylene, sintered stainless steel, sintered PTFE, among other options.

[0249] After traveling over the second encapsulation member 310 the fluid may then travel to the outer channel 212 formed by the void between the exterior surface of the separation media 312 and the interior radial wall 206 of the capsule body 200. The fluid may begin at the second end of the outer channel 212B and portions thereof may travel towards the first end of the outer channel 212A while simultaneously other portions may travel radially inward through separation media 312. Such portions of the fluid pass through the separation media 312 and into the inner channel 214 which is adjacent to an interior surface of the separation media 312 and the core 600. The fluid may then travel down the inner channel 214 around the core 600 to the underside of the first encapsulation member 302. As fluid travels between an end of the core 600 and the lid 100, all the fluid will pass through the second frit 850. The second frit 850 can help with retention of the stationary phase. Further, the second frit 850 can assist with ensuring uniform and consistent flow. After passing through the second frit 850, the fluid will travel down and out of the first port 102.

[0250] Should the second frit 850 be at a beginning of the fluid flow such as would be the case for Figure 11A (should Figure 11A include a second frit 850), the second frit 850 can perform each of the functions that the first frit 800 assisted with mentioned above such as acting as a filter.

[0251] As can be seen in Figures 11A-13D, the first connector 402, 1402 along with the first port 102, 1102 is always associated with the lid 100, 1100, and the second connector 204, 1204 and second port 202, 1202 are always associated with the capsule body 300, while flow may be reversed or inverted within a system. Said system can include a chromatography work station, an FPLC, an Akta system, a process chromatography skid, a membrane chromatography skid, etc.

[0252] As shown in Figure 11F the first connector 402, 1402 and the second connector 204, 1204 (including the first port 102, 1102 and the second port 202, 1202 respectively) can be permanently attached to either of the lid 100, 1100, or the capsule body 200, 1200 and need not be modularized, or in the alternative can be modularized as shown in Figures 11 A, HE. In the permanently attached scenario shown in Figure HF, the first connector 402 and the second connector 204 are capable of connecting to the same connections of a system such that an entirety of the device 000 is provided in reverse orientation. Such modularization and ability to invert, or operate in reverse, the separation device 000, 1000 makes for convenience in some situations, such as greater flexibility in how the separation device 000, 1000 connects to a chromatography work station, an FPLC, an Akta system, a process chromatography skid, a membrane chromatography skid, etc., while having them permanently attached makes for convenience in other situations such as simplification in manufacturing and assembly. Such features are not limiting unless expressly claimed.

[0253] One or more instruments can be utilized for identifying and / or quantifying the presence or absence of particular materials in the effluent (including, but not limited to, the target component). Preferably, any instruments are in fluid communication with the separation device(s) such an instrument is present before and / or after the separation device. Instruments can include, but are not limited to, a chromatography workstation, a fast protein liquid chromatography system (“FPLC”), an Akta system, a process chromatography skid, a membrane chromatography skid, a high performance liquid chromatography instrument (“HPLC”), an ultra-performance liquid chromatography instrument (“UPLC”), a next generation chromatograph (“NGC”), a mass chromatograph (“MS”), a gas chromatograph(“GC”), a colorimeter, a mass spectrometer, a fluorometer, a photometer, a spectrometer, a spectrophotometer, a X-ray photoelectron spectrometer (“XPS”), a sequencing instrument, a next generation sequencing (“NGS”) sequencing instrument, a UV detector, and a visible light detector,.

[0254] Shown in Figure 1 IF is a sensor 999 which can be inserted at an upstream end so as to monitor fluid entering the second port 202. The sensor 999 can in the alternative be placed at a downstream end so as to monitor fluid exiting the first port 102. The sensor may be inserted via either of the first connector 402 or the second connector 204. This is not limiting as to where the sensor 999 may be placed as the sensor 999 can be placed anywhere on and inserted anywhere into the separation device 000. The sensor 999 may further comprise a plurality of sensors 999. The plurality of sensors 999 may thus include at least 1 sensor 999, at least 2 sensors 999, at least 3 sensors 999, at least 4 sensors 999, at least 5 sensors 999, at least 6 sensors 999, at least 7 sensors 999, at least 8 sensors 999, at least 9 sensors 999, at least 10 sensors 999, etc. Furthermore, the plurality of sensors 999 may comprise different sensors in combination. For example, the plurality of sensors 999 may include a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector, a diode array detector, or any combination thereof. Any or all of which may optionally be connected to a recording device. The sensor 999 is not limited to the embodiment shown in Figure HF. The sensor 999 and / or the plurality of sensors 999 can be included in any of the embodiments shown in Figures 11 A, HE, 12A-12D, 13A-13D in addition to that shown in Figure HF.Furthermore, the sensor 999 or the plurality of sensors 999 can be attached to and / or inserted into flow lines 702, 704 described infra.

[0255] Figures 12A-12D will be discussed together. As shown in Figure 12B, a separation device 1000 comprises four main components: a lid 1100, a capsule body 1200, a separation media assembly 1300, and a bleed-valve 1450. In a preferred embodiment, the separation media assembly 1300 is housed at least partially within both the lid 1100 and the capsule body 1200 as shown in Figure 12A.

[0256] The separation media assembly 1300, shown in greater detail in Figure 12B, comprises a separation media 1312, a first encapsulation member 1302, and optionally, but preferably, a second encapsulation member 1310. The separation media 1312 is pleated(although it could be rolled). As shown in Figure 12D, an interior of the pleats 1313 can be seen.

[0257] The first and second encapsulation members 1302, 1310, of the separation media assembly 1300 are preferably formed via a chemical elastomeric encapsulant, such as a silicone elastomer or a polyurethane elastomer, which forms and accommodates the geometries of the separation media sealing interfaces, in addition to hermetically sealing the separation media 1312. Preferably, the chemical elastomeric encapsulate has some degree of malleability and deformability. Beneficially, in preferred embodiments, the first and second encapsulation members 1302, 1310 seal off portions of the separation media which are encapsulated thereby reducing dead volume and forcing fluid flow through the desired portion of the separation media.

[0258] The first encapsulation member 1310 can comprise flow directing features such as channels 1316 (or tapering) to provide for more efficient flow of fluid. The channels 1316 are preferably grooves that the fluid can flow through thereby having a larger fluid path such that fluid flows through the desired flow path and the device can be operated at the desired pressure and speed.

[0259] The lid 1100 includes a second port 1202 which may be disposed within a second connector 1204, and the second connector 1204 may be included as a part of the lid 1100 or alternatively exist as a modular component that may detach and attach to the lid 1100. In some embodiments, such as those shown in Figures 12B, 12D, the lid 1100 also includes a lid extension 1114 on an underside of the lid 1100. The lid extension 1114 can aid in securing the capsule body 1200 by resisting upward motion being snapped in place or sonic welded to an overhang 1116 which facilitates a tighter seal from fluid leakage. Interlocking features of the overhang 1116 of the lid 1100 interacting with the capsule body 1200, and more specifically a flange 1220, help secure the lid 1100 to the capsule body 1200. Such features are non-limiting unless expressly claimed.

[0260] The capsule body 1200 can include a wall 1224 that is substantially flush with an interior of the lid extension 1114 when placed therein. As shown in Figure 12D, the wall 1224 may include a ridge 1210 that interacts with the first encapsulation member 1302 to seal a volume interior to the wall 1224 and exterior to the separation media 1312 such that fluid flowing through the separation device 1000 will not enter an area between the lid extension 1114 and the wall 1224. Further, as shown in Figure 12D the first encapsulation member 1302may include ridges 1304 that also assist in sealing. The ridges 1304 may alternatively exist on the lid 1100 to interface with the first encapsulation member 1302 instead of the ridges 1304 existing on the first encapsulation member 1302 which would simply be opposite an arrangement shown in Figure 12D, and likewise ridges 1210 may alternatively exist on the first encapsulation member 1302 to interface and seal with the wall 1224 instead of the ridges 1210 extending from the wall 1224. Features such as the ridges 1210, 1304 are non-limiting unless expressly claimed.

[0261] The first port 1102 extends through the lid 1100 as well as a first connector 1402 that attaches to the lid 1100.

[0262] In the capsule body 1200 embodiment of Figures 12A-12D, the bleed-valve 1450 may be included. The bleed-valve 1450 includes a screw 1452 and bleed-valve aperture 1222 extending through a wall 1208 as shown in Figure 12B. The bleed-valve 1450 allows for releasing of any air that may get caught within the separation device 1000. This is advantageous as a user may find small amounts of air enter the device 1000 when connecting it to a system such as to a chromatography work station, an FPLC, an Akta system, a process chromatography skid, a membrane chromatography skid, etc. The bleed-valve 1450 makes such transitions possible while maintaining the ability to have smooth fluid flow and removal of any blockages due to trespassing air.

[0263] As shown in Figure 12D, a fluid enters the separation device 1000 via the second port 1202 and fluid diverges upon coming in contact with the second encapsulation member 1310 to travel around the second encapsulation member 1310. As shown in Figure 12D, the flow path is the same as that shown in Figure HE. The fluid coming in contact with the second encapsulation member 1310 after entering the second port 1202 travels radially outward across a surface of the second encapsulation member 1310. The second encapsulation member 1310 could include ridges such as that shown in Figure HE to guide the fluid, or could alternatively be shaped as a dome. The dome could range from that of a shallow dome (barely noticeable that the second encapsulation member 1310 is not flat) to that of a halfcircle, wherein the fluid may spread evenly over a surface of the dome due to its symmetry. In yet other embodiments, the surface of the second encapsulation member 1310 could be neither of a dome or include any ridges, but instead be flat and include a frit as shown in the embodiment of Figure HF. Such features are non-limiting unless expressly claimed.

[0264] After traveling over the second encapsulation member 1310 the fluid may then travel to the outer channel 1212 formed by a void between the exterior surface of the separation media 1312 and the interior radial wall 1206 of the capsule body 1200. After entering the void between the exterior surface of the separation media 1312 and the interior radial wall 1206, fluid passes through the separation media 1312 and into the inner channel 1214 which is between an interior surface of the separation media 1312 and the core 1600. The fluid may then travel down the inner channel 1214 around the core 1600 to an underside of the first encapsulation member 1302. As fluid travels between an end of the core 1600 and the lid 1100, the fluid will then travel down and out of the first port 1102. As shown in Figure 12D, the first connector 1402 and a second connector 1204 may optionally be detachable and thus modularized for adaptability in connectivity and flexibility with differing chromatography systems.

[0265] Note for Figures 12A-12D that a center void 1320 is not shown as the separation media assembly 1300 is already assembled, but the center void 1320 occupies a space similar to that shown in Figure 10C, wherein the core 1600 may reside within the center void 1320 as shown with the void 320 and the core 600 shown in Figure 10D.

[0266] Figures 13A-13D will be discussed together. As shown in Figure 13B, a separation device 2000 includes a number of components: a lid 2100, a sealing lid 2150, a capsule body 2200, a second connector 2204, a sealing body 2250, a handle 2275, a separation media assembly 2300, a first connector 2402, a bleed-valve 2450, and a conduit 2900. In a preferred embodiment, the separation media assembly 2300 is housed at least partially within both the lid 2100 and the capsule body 2200 as shown in Figure 13A, and the lid 2100 and the capsule body 2200 are housed at least partially within both the sealing lid 2150 and the sealing body 2250.

[0267] The separation media assembly 2300, shown in greater detail in Figure 13B, comprises a separation media 2312, a first encapsulation member 2302, and optionally, but preferably, a second encapsulation member 2310. The separation media 2312 is pleated (although it can be rolled). As shown in Figure 13D, an interior of the pleats 2313 can be seen beneath a core 2600 of the separation media assembly 2300.

[0268] The first and second encapsulation members 2302, 2310, of the separation media assembly 2300 are preferably formed via a chemical elastomeric encapsulant, such as a silicone elastomer or a polyurethane elastomer, which forms and accommodates thegeometries of the separation media sealing interfaces, in addition to hermetically sealing the separation media 2312. Preferably, the chemical elastomeric encapsulate has some degree of malleability and deformability. Beneficially, in preferred embodiments, the first and second encapsulation members 2302, 2310 seal off portions of the separation media which are encapsulated thereby reducing dead volume and forcing fluid flow through the desired portion of the separation media 2312.

[0269] The first encapsulation member 2310 can comprise flow directing features such as channels 2316 (or tapering) to provide for more efficient flow of fluid. The channels 2316 are preferably grooves that the fluid can flow through thereby having a larger fluid path such that desired flow path is followed and the device can be operated at the desired pressure and speed.

[0270] The separation media assembly 2300 can be housed at least partially within the capsule body 2200. The capsule body 2200 may include a radial wall 2206 that further includes a slot 2209 to guide and support the conduit 2900.

[0271] As opposed to previous embodiments, the first port 2102 extending through the lid 2100 does not travel solely straight. As shown in Figure 13D, the first port 2102 extends through the conduit 2900 such that fluid exiting the first port 2102 at the first connector 2402 is disposed on a same side of separation device 2000 as the second connector 2204. The first port 2102 includes a flow path exiting the lid 2100, entering the conduit 2900, and exiting the first connector 2402, and the first port 2102 includes a flow path entering the second connector 2204, the sealing body 2250, the capsule body 2200, each of which are called out in Figure 13B.

[0272] The lid 2100 as shown in Figure 13B includes an aperture 2118 disposed within a protrusion 2120 which each will be discussed more further below. The lid 2100 further includes a flange 2122 to assist with connection to the sealing lid 2150. The flange 2122 may assist with connection to the sealing lid 2150 in that a portion of the lid 2100 beneath the flange 2122 is shaped to snuggly fit within a ring 2152 protruding upward from the sealing lid 2150. The lid 2100 may be press-fitted into the sealing lid 2150, shrink fitted or thermal expansion fitted together, or sonic welded to the sealing lid 2150. Furthermore, less permanent embodiments of attaching are possible such that the lid 2100 is simply placed within the sealing lid 2150 for a snug fit, or the lid 2100 can be threaded and an interior of the ring 2152 can also be threaded to allow for a threaded twisting connection between the lid 2100 and the sealing lid 2150.

[0273] The lid 2100 further includes a plurality of connection points 2124 to facilitate attaching to the capsule body 2200. The connection points 2124 can be apertures to allow for screws, bolts, etc. to enter therethrough and fixedly attach to the capsule body 2200. Alternatively, the plurality of connection points 2124 can be snapping connections in which the connection points 2124 would extend upward as T-shaped or mushroom-shaped protrusions to snap into the capsule body 2200.

[0274] Further, the lid 2100 includes a side aperture 2126, which may be utilized in conjunction with the aperture 2118 mentioned earlier by each connecting to the conduit 2900. These components assist with connection of the conduit 2900 to the separation device 2000, and can most clearly be seen (how they assemble together) in Figure 13D. The conduit 2900 and the aperture 2118 may each include the first port 2102 contained within. A first end 2902 of the conduit 2900 may be inserted into the aperture 2118, while a main body of the conduit 2900 may extend through the side aperture 2126. The conduit 2900 may include a second end 2904 which corresponds to the first connector 2402. The conduit 2900 may further include a first bend 2906 and a second bend 2908 which may each be substantially perpendicular. While perpendicularity is not required, including two perpendicular bends facilitates for ease in getting the flow path to turn 180-degrees such that the first connector 2402 and the second connector 2204 extend substantially a same direction away from the separation device 2000. Should the first bend 2906 or the second bend 2908 be at an acute angle while the other be at an obtuse angle, this can impede flow at the acute angle and thus may require additional bends in the conduit 2900 to achieve substantially a 180-degree turn about of the flow path to avoid acute angles in changing flow in order to reduce an amount of turbulent flow and / or backflow. Further, rather than including the first bend 2906 and the second bend 2908, the conduit 2900 can include a single U-bend such that the conduit 2900 makes a 180-tum of the flow path in the shape of a “U”, wherein a first side of the “U” is shorter than a second side of the “U”, the first end 2902 of the conduit 2900 corresponding to the shorter and the second end 2904 corresponding to a longer side of the “U”. In this regard, the conduit 2900 may accomplish substantially a 180-degree turn of the flow path in a gradual or gentle manner. In this regard, the conduit 2900 redirects flow substantially 180-degrees with respect to flow that enters the first port 2102 at the aperture 2118. “Substantially 180-degrees” in this context means redirecting flow to anything between 135-225 degrees, although a perfect 180-degree redirection of flow may be preferred. It should be understood that various designs can beutilized to achieve having the connectors on the same side or in locations that are not directly opposed to each other; such can be through a conduit as depicted herein or otherwise through other designs such as one or more separate conduits or one or more integral (i.e. built-in) channels or ports.

[0275] As shown in Figure 13D, the lid 2100 accommodates a shape of the conduit 2900 for the conduit 2900 to travel therethrough with a left side of Figure 13D showing a thick portion and a right side of Figure 13D showing a hollowed-out portion for the conduit 2900 to reside within. The lid 2100 may accommodate the shape of the conduit 2900 whatever the shape of the conduit 2900 may be (e.g., two substantially perpendicular bends as shown, or a “U” shape as described above, etc.). Further, this may include not having a hollowed-out portion at all or even a side aperture 2126, such that the conduit 2900 extends upward (with reference to Figure 13D) outside of the lid 2100. As shown in Figure 13D, the conduit extends through the side aperture 2126 for greater stability for the conduit 2900. The conduit 2900 may be incorporated into previous embodiments herein described such as the separation device 000 and the separation device 1000. Such features describing the lid 2100 and the conduit 2900 are nonlimiting unless expressly claimed.

[0276] Including the conduit 2900 is advantageous to typical chromatography set-ups in that including both the first connector 2402 and the second connector 2204 on a same side of the separation device simplifies installation and maintenance. Given the modularity of the separation device 000, 1000, 2000, removing a single separation device from a chromatography set-up often results in unintentional spilling of fluid if the first connector 402, 1402, 2402 and the second connector 204, 1204, 2204 are on opposite sides because gravity makes fluid spill either from the system as the single separation device is removed, or from the single separation device no matter the orientation of the single separation device. However, when both the first connector 402, 1402, 2402 and the second connector 204, 1204, 2204 are on the same side of the separation device, and optionally positioned on the same plane, this allows for a more controlled approach to keeping fluid contained within the single separation device without leaking.

[0277] The lid 2100 further includes the protrusion 2120 as mentioned earlier to facilitate with connecting the lid 2100 to separation media assembly 2300. As shown in Figure 13D, the first encapsulation member 2302 of the separation media assembly 2300 may include ridges 2304 that engage the protrusion 2120 of the lid 2100. In this regard, the ridges 2304 seal aninterfacing between the protrusion 2120 and the first encapsulation member 2302 such that fluid or gas may not travel therebetween.

[0278] In the embodiment of Figures 13A-13D, the separation device 2000 may include the capsule body 2200. The capsule body 2200 may include a bleed-valve aperture 2222 disposed on a wall 2225 that is substantially circular and may include a wall 2206 extending substantially perpendicular to the wall 2225 around a perimeter of the wall 2225. The capsule body is configured to house the separation media assembly 2300. The capsule body may further include the slot 2209 to guide and secure the conduit 2900. Further, as seen in Figure 13D, the capsule body 2200 may include a ridge 2210 extending from a distal end of the wall 2206 which aids in sealing the separation device 2000. Preferably, the ridge 2210 may comprise a plurality of ridges 2210 to further assure sealing of the separation device 2000 from leaks or intrusion of foreign substances.

[0279] In the embodiment of Figures 13A-13D, the separation device 2000 may include the sealing body 2250. The sealing body 2250 encompasses at least in part the capsule body 2200 when the separation device 2000 is assembled together and may connect to the sealing lid 2150. Non-limiting examples of connecting the sealing lid 2150 and the sealing body 2250 include connecting by being sonic welded to each other, or snap-fitted, or threadedly attached among options known to those of ordinary skill in the art. In the embodiment of Figures 13A-13D a cross-section of the ring 2152 can be seen to comprise an upside-down L-shape. This shape as shown in Figure 13D can assist in snapping to or assist in the sonic welding of the sealing lid 2150 to a wall 2254 of the sealing body 2250. The sealing body 2250 can aid in further securing the lid 2100, the capsule body 2200, the separation media assembly 2300, and the conduit 2900 all together in proper alignment, as well as securing these components from outside interferences such as foreign objects. As shown in Figure 13B, the sealing body 2250 further includes handles 2275 that can be connected thereto via handle apertures 2280 for attaching the handles 2275 to the sealing body 2250. In some examples, the handle apertures 2280 are bare (not threaded) and are thus configured to allows a screw or a bolt to pass through and then threadedly connect to the handles 2275. In yet other examples, the handles 2275 may include protrusions spaced apart the same as the handle apertures 2280 so as to be inserted and either be snap connected or sonic welded together. The handles 2275 provide not only increased stability as gripping portions for a user, but also provide protection for each of the first connector 2402, the second connector 2204, and the bleed-valve 2450 by guarding againstimpact or contact of foreign objects to these components atop the sealing body 2250 on a wall 2252. In some examples as shown in Figures 13C-13D, the handles are substantially as tall as the first connector 2402 and the second connector 2204. Features such as the handles 2275 and apertures are not limiting unless expressly claimed.

[0280] Further, the sealing body 2250 may include a conduit aperture 2290. The conduit aperture 2290 is configured to guide and secure the conduit 2900 to exit the separation device 2000 and provide a place for the conduit 2900 to reside. The sealing body 2250 may include the conduit aperture 2290 on a side of the sealing body 2250 rather than atop the sealing body 2290 (as seen in Figure 13B). Further, the conduit 2900 may simply extend out of the separation device 2000 such that no conduit aperture 2290 is needed. In a preferred embodiment, the conduit aperture 2290 is disposed atop the sealing body 2250 such that the conduit 2900 is secured within the sealing body 2250 to reduce unwanted interference, contact, or impact from foreign objects.

[0281] In the embodiment of Figures 13A-13D, the bleed-valve 2450 may be included. The bleed-valve 2450 includes a screw 2452, the first bleed-valve aperture 2222 disposed on the wall 2252, and a second bleed-valve aperture 2223 disposed on a wall 2225 as shown in Figures 13A-13B. Given the nature of the first bleed-valve aperture 2222 and the second bleedvalve aperture being aligned and utilizing the same screw 2452, they may be referred to as the bleed-valve apertures 2222, 2223. The bleed-valve 2450 allows for releasing of any air that may get caught within the separation device 2000. This is advantageous as a user may find small amounts of air enter the device 2000 when connecting it to a system via the first connection 2402 and the second connection 2204. The bleed-valve 2450 makes such transitions possible while maintaining the ability to have smooth fluid flow and removal of any blockages due to trespassing air.

[0282] Note for Figures 13A-13D that a center void 2320 is not shown as the separation media assembly 2300 is already assembled, but the center void 2320 occupies a space similar to that shown in Figure 10C, wherein the core 2600 may reside within the center void 2320 as shown with the void 320 and the core 600 shown in Figure 10D.

[0283] As shown in Figure 13D, a fluid enters the separation device 2000 via the second port 2202 and fluid diverges upon coming in contact with the second encapsulation member 2310 to travel around the second encapsulation member 2310. As shown in Figure 13D, the flow path is the same as that shown in Figure HE until coming to the conduit 2900. The fluidcoming in contact with the second encapsulation member 2310 after entering the second port 2202 travels radially outward across a surface of the second encapsulation member 2310. The second encapsulation member 2310 can include ridges such as that shown in Figure HE to guide the fluid, or could alternatively be shaped as a dome. The dome could range from that of a shallow dome (barely noticeable that the second encapsulation member 2310 is not flat) to that of a half-circle, wherein the fluid may spread evenly over a surface of the dome due to its symmetry. Further, the second encapsulation member 2310 may include neither or ridges nor a dome, but rather be flat and could include a first frit and a second frit as explained above relating to the embodiment shown in Figure HF. Such features are non-limiting unless expressly claimed.

[0284] After traveling over the second encapsulation member 2310 the fluid may then travel to the outer channel 2212 formed by a void between the exterior surface of the separation media 2312 and an interior of the radial wall 2206 of the capsule body 2200. After entering the void between the exterior surface of the separation media 2312 and the interior radial wall 2206, fluid passes through the separation media 2312 and into the inner channel 2214 which is adjacent to an interior surface of the separation media 2312 and the core 2600. The fluid may then travel down the inner channel 2214 around the core 2600 to an underside of the first encapsulation member 2302. As fluid travels between an end of the core 2600 and the lid 2100, the fluid will travel down and out of the first port 2102.

[0285] Figure 14A shows a diagram of the separation device 000, 1000, 2000 of Figures HA, HE, 11F, 12A-D, and / or 13A-13D, wherein the second connector 402, 1402, 2402 is connected to a downstream flow line 704 and the first connector 204, 1204, 2204 is connected to an upstream flow line 702. As shown in Figure 14A, the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 can be disposed at any two differing locations on a surface of the separation device 000, 1000, 2000. Further, Figure 14A is shown in a 2- dimensional plane, however the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 may be disposed in any two differing locations covering all three dimensions. Thus, although Figure 14A is shown in a 2-D plane, this is not limiting as to where the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 may extend from the surface of the separation device 000, 1000, 2000 so as to accommodate flow lines 702, 704 coming from any direction towards the separation device 000, 1000, 2000. In a preferred embodiment, the flow lines 702, 704 are parallel to one another which may mean that the upstream flow line 702 isdisposed on a side directly opposite the downstream flow line 704 such as that shown in Figure 14B, or this may mean that the upstream flow line 702 and the downstream flow line 704 each extend towards a common plane to connect with the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 as would be the case for the embodiment shown in Figures 13A-D, or alternatively this may mean that the parallel flow lines 702, 704 extend beside one another to different planes such that they connect to the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 which each emerge from any position around the separation device 000, 1000, 2000.

[0286] Figure 14B shows a diagram of the separation device 000, 1000, 2000 of Figures 11 A, HE, HF, 12A-D, and / or 13A-13D in a configuration opposite to Figure 14A, namely wherein the first connector 204, 1204, 2204 is connected to the downstream flow line 704 and the second connector 402, 1402, 2402 is connected to the upstream flow line 702. In other words, the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 can switch places such that the separation device 000, 1000, 2000 has flipped around such that flow through the separation device 000, 1000, 2000 is opposite that shown in Figure 14A. Further, as shown in Figure 14B, it is not relevant where or how the first connector 204, 1204, 2204 or the second connector 402, 1402, 2402 protrude from separation device 000, 1000, 2000 so as to satisfy connecting to the flow lines 702, 704. Rather, there are different advantages that come from certain configurations, such as organizational benefits of running multiple separations devices 000, 1000, 2000 in a row in series would benefit most from having the flow lines 702, 704 parallel and attaching to the separation device 000, 1000, 2000 at positions directly opposite one another. Or, in another embodiment such as that shown in Figures 13A-13D, having the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 extending from a similar plane to meet the flow lines 702, 704 on a same side provides the benefit of not having fluid spill out of the separation device 000, 1000, 2000 when disconnecting from the flow lines 702, 704. Or, in another embodiment the first connector 204, 1204, 2204 and the second connector 402, 1402, 2402 extending from seemingly random positions from the separation device 000, 1000, 2000 and perhaps even bending, as shown in Figure 14B, could provide a benefit of simplest, quickest assembly and cheapest manufacturing process.

[0287] Furthermore, the flow lines 702, 704 shown in Figures 14A-14B are shown with break symbols as what the flow lines 702, 704 are connected to will vary from application toapplication. The upstream flow line 702 may come directly from (and connect to) a feed source of fluid to be eluted or otherwise treated, or alternatively the upstream flow line 702 may come from (and connect to) a filter source, or alternatively still, the upstream flow line 702 may come from and additionally be the downstream flow line 704 of another separation device 000, 1000, 2000. As such, the separation device 000, 1000, 2000 may comprise a series of separation devices 000, 1000, 2000 or separation devices 000, 1000, 2000 running in parallel. Multiple separation devices 000, 1000, 2000 can be incorporated into systems or kits by connecting separation devices 000, 1000, 2000 together in series or in parallel. Such patterns of connecting the separation devices 000, 1000, 2000 together in series or in parallel can be a part of a chromatography work station, an FPLC, an Akta system, a process chromatography skid, a membrane chromatography skid, etc. The system shown in Figures 14A-14B preferably further comprise one or more of a pump, a process controller, a fraction collector, and optionally one or more instruments and / or sensors.

[0288] Figure 14C provides a top perspective view of a non-limiting example separation device 000 having fluid lines attached both to the first connector 402 and the second connector 204, such that it is a system having an upstream flow line 702 in fluid communication with the first port 102. The upstream flow line 702 provides a crude feed comprising one or more target components in a gas or liquid media. The gas or liquid media then travels through the separation device 000 (as described further herein) and out the second port 202. The second port 202 is in fluid communication with a downstream flow line 704 through which the effluent passes through to any downstream applications. Downstream applications can include, but are not limited to, an instrument and / or sensor. In a preferred embodiment, the effluent is collected in a fraction collector. The system shown in Figure 14C preferably further comprises one or more of a pump, a process controller, a fraction collector, and optionally one or more instruments and / or sensors.

[0289] One or more instruments can be utilized for identifying and / or quantifying the presence or absence of particular materials in the effluent (including, but not limited to, the target component). Preferably, any instruments are in fluid communication with the separation device(s) such an instrument is present before and / or after the separation device. Instruments can include, but are not limited to, a chromatography workstation, a fast protein liquid chromatography system (“FPLC”), an Akta system, a process chromatography skid, a membrane chromatography skid, a high performance liquid chromatography instrument(“HPLC”), an ultra-performance liquid chromatography instrument (“UPLC”), a next generation chromatograph (“NGC”), a mass chromatograph (“MS”), a gas chromatograph (“GC”), a colorimeter, a mass spectrometer, a fluorometer, a photometer, a spectrometer, a spectrophotometer, a X-ray photoelectron spectrometer (“XPS”), a sequencing instrument, and a next generation sequencing (“NGS”) sequencing instrument. Preferred sensors can include a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector, a diode array detector, or any combination thereof, any or all of which may optionally be connected to a recording device. Preferred instruments and sensors are non-destructive.

[0290] The instruments and / or sensors can be in communication with a system control that detects a particular change state (e.g., presence or absence of a material in the effluent, change in pH, change in conductivity, etc.), such that the system control can stop the flow of crude feed, start a buffer cycle and / or a clean cycle (such as a clean-in-place).

[0291] Any number of separation devices 000, 1000, 2000 can be arranged in a system. Preferably a system comprises at least 1 separation device, at least 2 separation devices, at least 3 separation devices, at least 4 separation devices, or at least 5 separation devices; where the separation devices are in operation in parallel, series or a combination thereof. In an embodiment, a system comprises between 1 and 100 separation devices in series, parallel or a combination thereof.

[0292] In a system the separation devices can be arranged in series and / or in parallel. If arranged in parallel, the system has the ability to perform a separation on significantly more fluid over a period of time due to the increased volume achieved by use of separation devices 000, 1000, 2000. If arranged in a series, the system has the ability to employ different separation techniques and target different biological or non-biological substances as fluid flows through each separation device. In a preferred embodiment, the system comprising one or more separation devices can be arranged so that flow can be redirected from a first separation device to second separation device without a substantial break, or any break, in the flow while the first separation device is repaired, replaced, inspected, or otherwise removed from the fluid flow. The arrangement of the one or more separation devices in a system can be varied to suit the particular arrangement desired.

[0293] A preferred method of use comprises a rapid cycling mode, where the separation media is optionally prepared for a separation via a buffer cycle (where an equilibrium bufferis first passed through the separation device 000, 1000, 2000 via the first port 102, 1102, 2102 and out the second port 202, 1202, 2202); then crude feed is passed through an upstream flow line 702 into the separation device 000, 1000, 2000 via the first port 102 and out the second port 202 to the downstream flow line 704. After passing through the separation media 312 the crude feed is considered an effluent. The effluent is then preferably provided to an instrument and / or sensor via the discharge feed and then to a fraction collector. When a particular state is reached (whether by time or sensing of the presence or absence of a condition), the crude feed is no longer fed through the upstream flow line 702; optionally a purge cycle can be performed by removing any impurities and contaminants from the separation device 000, 1000, 2000 including the separation media 312; whether a purge cycle is performed or not, the target compound is then released from the separation media 312 via an elution cycle; preferably another buffer and / or clean cycle is performed to release any residual target compound; the same or another buffer cycle can be performed to regenerate the separation media 312 so that it is prepared for another separation cycle for capturing target components. These methods can be performed manually with a person overseeing the various steps, via a programmed method, or an automated system.

[0294] Accordingly, a system for separating one or more target molecules can comprise an upstream flow line 702, a separation device 000, a downstream flow line 704, optionally one or more instruments and / or sensors, and a fraction collector. The separation device 000 can comprise a capsule body 200 having a second port 202, a separation media assembly 300 located at least partially within the capsule body 200 and having a center void 320, a core 600 positioned in the center void 320 of the separation media 312, and a lid 100 having a first port 102; wherein the separation media assembly 300 can comprise a separation media 312 hermetically sealed within the capsule body 200, a first encapsulation member 302, and a second encapsulation member 310; wherein the separation media 312 can comprise one or more surface functionalizations; and wherein the separation device 000 is characterised in that fluid that flows between the first port 102 and second port 202 passes through the separation media 312 in a direction radial to a direction in which fluid passes through the first port 102 and / or second port 202. The upstream flow line 702 is in fluid communication with the first port 102, and the downstream flow line 704 is in fluid communication with the second port 202. Related methods of controlling fluid through the system and methods of separating one or more target components can be performed with this system as disclosed herein.

[0295] Beneficially, the separation device 000, 1000, 2000 disclosed herein is capable of rapid cycling, high flowthrough rates, while maintaining low pressure and minimal dead volume. The lack of pressure can aid in preventing damage to sensitive target compounds due to high pressure; further the design provides for limited sheer force on the target compounds; furthermore, the lack of pressure required can aid in reducing energy consumption of systems intended to capture and / or purify target compounds which otherwise may require pumps or other devices and / or methods capable of inducing high-pressure environments.

[0296] It should be understood that while operating at a higher flow rate can be preferable, the flow rate can be determined by the nature of the system providing fluid pressure, which can be limiting on the flow rate achieved. The separation device disclosed herein can provide high flow rates without failure or leaking. Preferably the flow rate of the fluid through the separation device is between about 5 MV / min and about 400 MV / min, more preferably between about 10 MV / min and about 300 MV / min, more preferably between about 15 MV / min and about 200 MV / min, most preferably between about 20 MV / min and about 100 MV / min. Preferably, the flow rate is at least about 5 MV / min, at least about 6 MV / min, at least about 7 MV / min, at least about 8 MV / min, at least about 9 MV / min, at least about 10 MV / min, at least about 11 MV / min, at least about 12 MV / min, at least about 13 MV / min, at least about 14 MV / min, at least about 15 MV / min, at least about 16 MV / min, at least about 17 MV / min, at least about 18 MV / min, at least about 19 MV / min, at least about 20 MV / min, at least about 21 MV / min, at least about 22 MV / min, at least about 23 MV / min, at least about 24 MV / min, at least about 25 MV / min, at least about 26 MV / min, at least about 27 MV / min, at least about 28 MV / min, at least about 29 MV / min, or at least about 30 MV / min. Preferably the flow rate is at or below about 400 MV / min, at or below about 375 MV / min, at or below about 350 MV / min, at or below about 325 MV / min, at or below about 300 MV / min, at or below about 275 MV / min, at or below about 250 MV / min, at or below about 225 MV / min, at or below about 200 MV / min, at or below about 190 MV / min, at or below about 180 MV / min, at or below about 170 MV / min, at or below about 160 MV / min, at or below about 150 MV / min, at or below about 140 MV / min, at or below about 130 MV / min, at or below about 120 MV / min, at or below about 110 MV / min, at or below about 100 MV / min, at or below about 95 MV / min, at or below about 90 MV / min, at or below about 85 MV / min, at or below about 80 MV / min, at or below about 75 MV / min, at or below about 70 MV / min, at or below about 65 MV / min, at or below about 60 MV / min, at or below about 55 MV / min, at or belowabout 50 MV / min, at or below about 49 MV / rnin, at or below about 48 MV / rnin, at or below about 47 MV / min, at or below about 46 MV / min, at or below about 45 MV / min, at or below about 44 MV / min, at or below about 43 MV / min, at or below about 42 MV / min, at or below about 41 MV / min, at or below about 40 MV / min, at or below about 39 MV / min, at or below about 38 MV / min, at or below about 37 MV / min, at or below about 37 MV / min, at or below about 36 MV / min, or at or below about 35 MV / min.

[0297] Preferably, during a separation method, the fluid flow is performed at a pressure of about 10 bar or less, about 9.5 bar or less, about 9.0 bar or less, about 8.5 bar or less, about 8.0 bar or less, about 7.5 bar or less, about 7.0 bar or less, about 6.5 bar or less, about 6.0 bar or less, about 5.5 bar or less, about 5.0 bar or less, about 4.5 bar or less, about 4.0 bar or less, about 3.5 bar or less, about 3.0 bar or less, about 2.9 bar or less, about 2.8 bar or less, about 2.7 bar or less, about 2.6 bar or less, about 2.5 bar or less, about 2.4 bar or less, about 2.3 bar or less, about 2.2 bar or less, about 2.1 bar or less, about 2.0 bar or less, about 1.9 bar or less, about 1.8 bar or less, about 1.7 bar or less, about 1.6 bar or less, about 1.5 bar or less, about 1.4 bar or less, about 1.3 bar or less, about 1.2 bar or less, about 1.1 bar or less, about 1.0 bar or less, about 0.9 bar or less, about 0.8 bar or less, about 0.7 bar or less, about 0.6 bar or less, about 0.5 bar or less, about 0.4 bar or less, about 0.3 bar or less, and about 0.2 bar or less.

[0298] During a separation method, the residence time in the separation device is preferably between about 0.1 seconds and about 5 minutes, more preferably from about 0.5 seconds to about 4 minutes, still more preferably from about 1 second to about 3 minutes. Preferably, the residence time is less than about 5 minutes, less than about 4.5 minutes, less than about 4 minutes, less than about 3.5 minutes, less than about 3 minutes, less than about 2.5 minutes, less than about 2 minutes, less than about 90 seconds, less than about 80 seconds, less than about 70 seconds, less than about 1 minute, less than about 55 seconds, less than about 50 seconds, less than about 45 seconds, less than about 40 seconds, less than about 35 seconds, less than about 30 seconds, less than about 25 seconds, less than about 20 seconds, less than about 19 seconds, less than about 18 seconds, less than about 17 seconds, less than about 16 seconds, less than about 15 seconds, less than about 14 seconds, less than about 13 seconds, less than about 12 seconds, less than about 11 seconds, less than about 10 seconds, less than about 9 seconds, less than about 8 seconds, less than about 7 seconds, less than about 6 seconds, less than about 5 seconds, less than about 4 seconds, less than about 3 seconds, about 2.9 seconds or less, about 2.8 seconds or less, about 2.7 seconds or less, about 2.6seconds or less, about 2.5 seconds or less, about 2.4 seconds or less, about 2.3 seconds or less, about 2.2 seconds or less, about 2.1 seconds or less, about 2 second or less, about 1.9 seconds or less, about 1.8 seconds or less, about 1.7 seconds or less, about 1.6 seconds or less, about 1.5 seconds or less, about 1.4 seconds or less, about 1.3 seconds or less, about 1.2 seconds or less, about 1.1 seconds or less, or about 1 second or less. Preferably, the residence time is at least about 0.1 seconds, at least about 0.2 seconds, at least about 0.4 seconds, or at least about 0.5 seconds.

[0299] Preferably the separation techniques employ fluid flow that is through micro- and macro-pores of the separation media (as opposed to tightly packed resin beads). In a preferred embodiment, the separation comprises adsorption occurring on the surface separation media, where no internal diffusion is required. In a preferred embodiment, there is minimal, more preferably no, high-pressure drops with elevated flow rates. This overcomes the difficulties faced with slow intra-particle diffusion required for adsorption within resin beads.

[0300] The separation device 000, 1000, 2000 can be manufactured such that it has a volume of from about one-half milliliter (0.5 mL) to about ten liters (10 L), more preferably from one-half milliliter (0.5 mL) to about five thousand milliliters (5000 mL), more preferably between one milliliter (1 mL) up to one liter (1 L), most preferably from about one milliliter (1 mL) to about five hundred milliliters (500 mL). In some embodiments, the separation device can be manufactured such that it has a volume of one milliliter (1 mL). In other embodiments, the separation device can be manufactured such that it has a volume of 10 milliliters (10 mL). In yet other embodiments still, the separation device can be manufactured such that it has a volume of 100 milliliters (100 mL).Example Embodiments

[0301] The inventions are defined in the claims. However, below is a non-exhaustive list of non-limiting embodiments in numbered format. Any one or more of the features of these embodiments may be combined with any one or more features of another example, embodiment, or aspect described herein. Accordingly, the following numbered clauses form part of the present disclosure but do not form part of the claims:

[0302] 1. A separation device (000, 1000, 2000) comprising: a capsule body (200); a separation media assembly (300) located at least partially within the capsule body (200); the separation media assembly (300) comprising a separation media (312) sealed within thecapsule body (200) and a first encapsulation member (302) and a second encapsulation member (310); wherein the separation media assembly (300) forms a center void (320); a core (600) positioned in the center void (320) of the separation media assembly (300); a lid (100); a first connector (402) proximate the lid (100) providing a first port (102) for fluid to pass through; a second connector (204) proximate the capsule body (200) providing a second port (202) for fluid to pass through; and characterised in that fluid can flow between the first connector (402) and the second connector (204) and can pass through the separation media (312) in a direction radial to a direction in which fluid passes through the second connector (204).

[0303] 2. The separation device (000) according to clause 1, wherein the lid (100) is friction-fitted, press-fitted, shrink-fitted, snap-fitted, sonic welded, welded, screwed, clamped, Luer-locked, glued, adhered or otherwise secured together and sealed to the capsule body.

[0304] 3. The separation device (000) according to clause 1, wherein the lid (100) further comprises a primary interface that hygienically couples a geometry of the first connector (402) with an inside region (306) of the separation media assembly (300).

[0305] 4. The separation device (000) according to clause 3, wherein the primary interface comprises a radial static seal (400) established with a connector (402) formed into the lid (100) and directly coupled to a sealing face (404) on an underside of the lid (100); preferably wherein the connector (402) comprises any of a Luer, a Luer-lock, a tri-clamp connection, a ferrule, and / or a threaded fitting.

[0306] 5. The separation device (000) according to clause 4, wherein a thread of the threaded fitting comprises any of a Unified national fine (UNF) thread, a G-thread, a national pipe thread (NPT), or a metric thread, or any other threading type.

[0307] 6. The separation device (000) according any one of clauses 1-5, wherein the first encapsulation member (302) is made of an elastomeric material and is hermetically sealed to the separation media (312).

[0308] 7. The separation device (000) according to clause 6, wherein the elastomeric material is a silicone elastomer, a polyurethane elastomer, or a combination thereof.

[0309] 8. The separation device (000) according to any one of clauses 1-7, wherein the second connector (204) is centrally located on the capsule body (200).

[0310] 9. The separation device (000) according to any one of clauses 1-8, wherein the separation media (312) comprises one or more surface functionalizations.

[0311] 10. The separation device (000) according to any one of clauses 1-9, wherein the first port (102) is centrally located on the lid (100).

[0312] 11. The separation device (000) according to any one of clauses 3 or 4, wherein the primary interface comprises a flange static seal (500) established by a connector (502) that is part of an insert (504) that interfaces with the lid (100).

[0313] 12. The separation device (000) according to clause 11, wherein the connector(502) comprises any of: a Luer, a Luer-lock, a tri-clamp connection, a ferrule, or a threaded fitting.

[0314] 13. The separation device (000) according to clause 12, wherein a thread of the threaded fitting comprises any of: a Unified national fine (UNF) thread, a G-thread, a national pipe thread (NPT), or a metric thread.

[0315] 14. The separation device (000) according to clause 13, wherein the insert (504) compresses a sealing ring (508) that is integral to a first encapsulation member (302) between an insert flange (506) and the lid (100).

[0316] 15. The separation device (000) according to clause 14, further comprising a retaining feature (510) that maintains compression between a retaining component (512) and the lid (100).

[0317] 16. The separation device (000) according to clause 15, wherein the retaining component (512) is threaded, grooved, or moulded in-place.

[0318] 17. The separation device (000) according to any one of clauses 1-16, wherein the first encapsulation member (302) has an extended portion (303).

[0319] 18. The separation device (000) according to clause 17, wherein the second encapsulation member is made of an elastomeric material and is hermetically sealed to the separation media (312).

[0320] 19. The separation device (000) according to clause 18, wherein the elastomeric material is a silicone elastomer, a polyurethane elastomer, or a combination thereof.

[0321] 20. The separation device (000) according to any one of clauses 3-19, wherein the lid (100) further comprises a secondary interface with the separation media assembly (300)that provides a hygienic seal between the lid (100) and the capsule body (200), and wherein the secondary interface comprises a ridge (104 / 210) that interacts with the first encapsulation member (302).

[0322] 21. The separation device (000) according to any one of clauses 1-20, wherein the capsule body (200) comprises a primary interface that hygienically couples the capsule body (200) with an outside region (308) of the separation media assembly (300).

[0323] 22. The separation device (000) according to any one of clauses 1-21, wherein the first port (102) is parallel with the second connector (204) and the flow is passed through the separation media (312) in a direction radial to a direction in which flow passes through the second connector (204).

[0324] 23. The separation device (000) according to any one of clauses 1-22, further comprising a connector (204) that is formed into the capsule body (200).

[0325] 24. The separation device (000) according to any one of clauses 1-23, wherein an interior radial wall (206) of the capsule body (200) and an exterior surface of the separation media (312) provide an outer channel (212) that tapers.

[0326] 25. The separation device (000) according to any one of clauses 1-24, wherein an interior surface of the separation media (206) of the capsule body (200) and the core (600) provide an inner channel (214) that tapers.

[0327] 26. The separation device (000) according to any one of clauses 1-25, wherein the outer channel (212) and the inner channel (214) taper at a slope inverse to each other.

[0328] 27. The separation device (000) according to any one of clauses 1-26, wherein the interior radial wall (206) interfaces with a second encapsulation member (310) such that the flow is permitted with a controlled resistance.

[0329] 28. The separation device (000) according to any one of clauses 1-27, wherein the first encapsulation member (302) and second encapsulation member (310) are made of the same elastomeric material.

[0330] 29. The separation device (000) according to clause 28, wherein the core (600) is made of the same elastomeric material.

[0331] 30. The separation device (000) according to any one of clauses 1-29, wherein the separation media (312) comprises a join (314) that is thermally bonded or encapsulated.

[0332] 31. The separation device (000) according to any one of clauses 1-30, wherein the capsule body (200) comprises internal ridges (218).

[0333] 32. The separation device (000) according to any one of clauses 1-31, wherein the core (600) maintains an equidistant flow-path reducing void volume inside the separation media (312).

[0334] 33. The separation device (000) according to any one of clauses 1-32, wherein the core (600) is hygienically coupled to a geometry of the first port (102) and tapered.

[0335] 34. The separation device (000) according to any one of clauses 1-33, wherein the core (600) further comprises one or more fins and / or vents.

[0336] 35. The separation device (000) according to any one of clauses 1-34, wherein a fluid enters the separation device (000) via a first connector and the fluid is directed by a diverter (602) to an interior surface of the separation media (312).

[0337] 36. The separation device (000) according to any one of clauses 1-35, wherein the separation media (312) comprises a hydrogel, one or more membranes, a monolith, or a matrix.

[0338] 37. The separation device (000) according to clause 36, wherein the separation media (312) comprises one or more fibrous membrane or a cast membrane.

[0339] 38. The separation device (000) according to clause 37, wherein the fibrous membrane comprises microfibers, nanofibers, or a mixture thereof.

[0340] 39. The separation device (000) according to clause 37 or 38 wherein the fibrous membrane is an electrospun membrane.

[0341] 40. The separation device (000) according to any one of clauses 1-39, wherein the membrane is pleated or rolled.

[0342] 41. The separation device (000) according to any one of clauses 37-40, wherein the separation media (312) comprises multiple layers, wherein the fibrous membrane comprises a primary functional layer (318A) attached to a secondary inert layer (318B).

[0343] 42. The separation device (000) according to any one of clauses 1-41, wherein the separation media comprises a polymer comprising collagen, chitosan, agarose, agarose acetate, cellulose, cellulose acetate, crosslinked cellulose, derivatized cellulose, regenerated cellulose, polyacrylic acid, polymethacrylic acid, polyacrylonitrile, a polyethylene oxide, a polyimide, a polyamide, polystyrene, a polysulfone, polycaprolactone, or a copolymer or mixture thereof.

[0344] 43. The separation device (000) according to any one of clauses 1-42, wherein the one or more surface functionalizations are in a density on the separation media of from about 1 mg / g to about 999 mg / g.

[0345] 44. The separation device (000) according to any one of clauses 1-44, wherein the surface functionalized separation media has a dynamic binding capacity on a volume basis of between about 1 pg / mL of the separation media and about 400 mg / mL of the separation media.

[0346] 45. The separation device (000) according to any one of clauses 1-44, wherein the one or more surface functionalizations comprise an ion exchange group, a polar group, a hydrophobic group, an affinity ligand, a mixed mode ligand, or a combination thereof.

[0347] 46. The separation device (000) according to any one of clauses 1-21, further comprising a conduit (2900) attached to the capsule body (200), and the second port (202) is attached to the lid via the conduit (2900) such that the conduit (2900) redirects flow substantially 180-degrees with respect to flow that enters the first port (102).

[0348] 47. The separation device (000) according to clause 46, wherein the first connector (2402) and the second connector (2204) are positioned on the same plane.

[0349] 48. The separation device (000) according to clause 47, wherein the lid (100) further comprises a side aperture (2126) through which the conduit (2900) passes through.

[0350] 49. The separation device (000) according to any one of clauses 1-48, further comprising a bleed-valve disposed on the capsule body (200).

[0351] 50. The separation device (000) according to clause 49, wherein the bleed-valve is adjacent to the second connector (204).

[0352] 51. The separation device (000) according to any one of clauses 1-50, further comprising a first frit (800) disposed between the lid (100) and the second encapsulation member (310).

[0353] 52. The separation device (000) according to any one of clauses 1-51, further comprising a second frit (850) disposed between the core (600) and the capsule body (200).

[0354] 53. The separation device (000) according to clause 44, wherein the one or more surface functionalizations comprise an anion exchange group, a cation exchange group, or a mixture thereof.

[0355] 54. The separation device (000) according to clause 53, wherein the ion exchange group comprises one or more of a tertiary amine, a quaternary amine, a quaternaryammonium, 6-chloranyl-3-[(2-pentyl-2,3-dihydro-l,3-thiazol-4-yl)methyl]quinazolin-4-one, diethyl-(2 -hydroxy-propyl) aminoethyl, diethylaminoethyl, trimethylaminoethyl, — N+(CH3)3, — N+(C2H5)H, — CH2CH2N+(CH3)3, — O— CH2CH2— N+(CH3)3, — CH2CH2N+(C2H5)H, — CH2CH2N+(C2H5)2(CH2CH(OH)CH3), — CH2CH2N+(CH3)2H, a carboxylate, a sulphonate, a phosphonate, — CH2COO“, — O-CH2COO“, — CH2OCH2COO“, — SO3“, — CH2CH2CH2SO3’, — CH2CH2SO3’, — P(OH)2O“, or a mixture thereof.

[0356] 55. The separation device (000) according to clause 54, wherein the one or more surface functionalizations comprise a hydrophobic group.

[0357] 56. The separation device (000) according to clause 55, wherein the hydrophobic group comprises a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, a hexyl group, an octyl group, a decyl group, a phenyl group, or a mixture thereof.

[0358] 57. The separation device (000) according to clause 45, wherein the one or more surface functionalizations comprise an affinity ligand.

[0359] 58. The separation device (000) according to clause 57, wherein the affinity ligand comprises a monoclonal antibody, a polyclonal antibody, an antibody fragment, a bacterial immunoglobulin binding protein, a chemical ligand, a dye, an enzyme inhibitor, histidine, an immobilized metal ion, a nucleic acid, an oligonucleotide, a lectin, a protein, an oligopeptide, a polysaccharide, an oligosaccharide, a sugar, a peptide, a polypeptide, an antigen, an aptamer, an affimer, an affibody, a small molecule biomimetic ligand, a small organic compound, a synthetic affinity ligand, a triazine ligand, or a mixture thereof.

[0360] 59. The separation device (000) according to clause 58, wherein affinity ligand is Protein A, Protein G, or Protein L.

[0361] 60. The separation device (000) according to clause 58, wherein the affinity ligand comprises IgG, IgM, a monoclonal antibody, a camelid antibody, an antibody fragment, Fab, Fv, or a mixture thereof.

[0362] 61. The separation device (000) according to clause 45, wherein the one or more surface functionalizations comprise a mixed mode ligand.

[0363] 62. The separation device (000) according to clause 61, wherein the mixed mode ligand comprises at least two of an anion exchange group, a cation exchange group, a polar group, a hydrophobic group, and a hydrogen bonding group.

[0364] 63. The separation device (000) according to clause 61 or 62, wherein the mixed mode ligand comprises an N-benzyl methyl ethanolamine group, an N-benzoyl-homocysteine group or mixture thereof.

[0365] 64. A method of controlling flow through the separation device (000) of according to any one of clauses 1-63, comprising: flowing a fluid through the lid (100) at the first connector (402); passing the fluid through the separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which fluid passes through the second connector (204); and passing the fluid through the second connector (204).

[0366] 65. The method according to clause 64, wherein the core (600) and / or a diverter (602) direct the flow to an inner channel (214) which tapers.

[0367] 66. The method according to any one of clauses 64-65, wherein the fluid exits the separation media (312) into an outer channel (212) which tapers.

[0368] 67. The method according to clause 66, wherein the outer channel (212) and the inner channel (214) taper inverse to each other.

[0369] 68. A method of controlling flow through the separation device (000) of according to any one of clauses 1-63, comprising: flowing a fluid through the capsule body (200) at the second connector (204); passing the fluid through separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the second connector (204); and passing the fluid through the second connector (204).

[0370] 69. The method according to clause 68, wherein the core (600) and / or a diverter (602) direct the flow to an inner channel (214) which tapers.

[0371] 70. The method according to any one of clauses 68-69, wherein the fluid exits the separation media (312) into an outer channel (212) which tapers.

[0372] 71. The method according to clause 69, wherein the outer channel (212) and the inner channel (214) taper inverse to each other.

[0373] 72. The method according to any one of clauses 63-70, wherein the fluid flow is laminar and an equivalent residence-time within the device is achieved by provisioning equidistant paths through the separation device (000).

[0374] 73. The method according to any one of clauses 63-71, further characterised by reversing the flow.

[0375] 74. A method of purifying a target component comprising: flowing a fluid through the first connector (402) or second connector (204) of the separation device according to any of clauses 1-63; passing the fluid through separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the second connector (204); separating the biomaterial from the fluid; and passing the fluid through the second port.

[0376] 75. The method of clause 74, wherein the target component comprises an amino acid, a biomolecule or fragment thereof, a peptide, an affimer, a protein, an enzyme, a glycoprotein, an LNP, a lipopolysaccharide, an antibody and / or a fragment thereof, a nucleic acid, an organic polymer, a virus, a VLP, an EV, an exosome, a bacterium, a cell, a cell- related structure, or a mixture thereof.

[0377] 76. The method of clause 75, wherein the antibody comprises a monoclonal antibody, a single-chain antibody, a bi-specific antibody, a multi-specific antibody, an antibody conjugate, an antibody fusion protein, a drug-antibody conjugate, an antibody fragment, a Fab fragment, a Fv fragment, a gamma globulin (IgG), an IVIG, an IgM, an IgA, an IgE, a hyperimmune gamma globulins, an isoagglutinin, or a combination thereof.

[0378] 77. The method of clause 75, wherein the virus comprises adeno-associated virus, lentivirus, or mixture thereof.

[0379] 78. The method of according to any one of clauses 64-77, wherein the fluid flow is performed at a pressure of about 1 bar or less.

[0380] 79. The method of according to clause 78, wherein the fluid flow is performed at a pressure of about 0.2 bar or less.

[0381] 80. The method according to any one of clauses 64-79, wherein the residence time is less than about 1 minute.

[0382] 81. The method according to clause 80, wherein the residence time is about 15 seconds or less.

[0383] 82. A system for separating one or more target molecules comprising:an upstream flow line 702, the separation device of any one of clauses 1-57, a downstream flow line 704, wherein the upstream flow line 702 is in fluid communication with the first port 102, and the downstream flow line 704 is in fluid communication with the second port 202.

[0384] 83. The system of clause 82, further comprising one or more instruments and / or sensors in fluid communication with the upstream flow line 702 and / or downstream flow line 704

[0385] 84. The system of clause 83, wherein the one or more instruments and / or sensors are a chromatography workstation, a fast protein liquid chromatography system (“FPLC”) an Akta system, a process chromatography skid, a membrane chromatography skid, a high performance liquid chromatography instrument (“HPLC”), an ultra-performance liquid chromatography instrument (“UPLC”), a next generation chromatograph (“NGC”), a mass chromatograph (“MS”), a gas chromatograph (“GC”), a colorimeter, a mass spectrometer, a fluorometer, a photometer, a spectrometer, a spectrophotometer, a X-ray photoelectron spectrometer (“XPS”), a sequencing instrument, and a next generation sequencing (“NGS”) sequencing instrument, a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector, a diode array detector, or any combination thereof, any or all of which may optionally be connected to a recording device.

[0386] 85. The system of any one of clauses 82-84, further comprising a fraction collector in fluid communication with the downstream flow line 704.

[0387] 86. The system of any one of clauses 82-85 wherein the system further comprises a pumping system such as a peristaltic pump, a diaphragm pump, a syringe pump, a positive displacement pump, a quattroflow pump, a centrifugal pump and a lobe pump connected in series and in fluid communication with a separation device and one or more instruments or sensors including, but not limited to, a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector or a diode array detector, or any combination thereof, any or all of which may optionally be connected to a recording device and / or a controller device.EXAMPLES

[0388] Preferred embodiments of the present disclosure are further exemplified in the following non-limiting Examples. It should be understood that these Examples, while indicating certain preferred embodiments, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the embodiments of the invention to adapt it to various usages and conditions. Thus, various modifications of the embodiments of the inventions, in addition to those shown and described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

[0389] Example 1. An experimental separation media was prepared in a pleated radial configuration. It comprised a fibrous membrane having weak ion exchange surface functionalization (DEAE as a non-limiting example), the membrane was pleated such that it had twelve pleats. The experimental separation media had both the primary functional layer (surface functionalized fibrous membrane) and a secondary nonfunctional layer, which was a spunbond nonwoven layer. The pleated membrane (1 ml membrane volume) was placed in a separation device (000) and secured with a lid; the first port and second port were established with fluid communication lines and the separation device (000) connected to a chromatography workstation equipped with UV and conductivity detectors. The separation device (000) was operated consistent with the flow path demonstrated in Figures 11A and 11C at a flow rate of 5ml / min. The separation device (000) was equilibrated with 20 mM Tris buffer, pH 8.0 and 82.5 ml Img / ml BSA solution (prepared in 20mM Tris buffer, pH 8.0) was loaded following which the separation device washed with 10ml of 20 mM Tris buffer, pH 8.0 and bound BSA eluted with 2M NaCl. Results from the experiment were recorded and are reflected in Figures 15-16

[0390] Figure 15 shows the breakthrough profile (A280) during BSA loading onto a separation device (000) fitted with 1 ml of DEAE functionalized, fibrous, pleated, membrane. The resulting breakthrough curve confirmed the liquid flow path is directed through the bed, and, importantly, not around it (z.e., the use of first and second encapsulation members 302, 310 worked to not only seal the join and ends of the fibrous membrane, but the first and second encapsulation members 302, 310 also acted as an efficient seal against the outer housing, thereby directing flow outwards, z.e., radially). This experiment also demonstrated a bindingcapacity of approximately 82.5 mg BSA / ml membrane volume with very little BSA breakthrough.

[0391] Figure 16 is a chromatograph of the BSA elution profile which indicates good chromatographic characteristics and a sharp elution peak, the majority of which was collected within 5 membrane volumes. Thus, the design of the separation device(s) described herein were found to excel in chromatographic performance and produce surprisingly good results.

[0392] Example 2. The BSA binding capacity and elution profile of a 1 ml membrane volume separation device of the type described in Example 1 was compared to 3 commercially available 1 ml anion-exchange membrane chromatography units. The membrane chromatography units were attached to a chromatography workstation equipped with a UV detector and equilibrated with 20mM Tris buffer, pH 8.0 at a flow rate of 5 ml / min. BSA solution, 2 mg / ml prepared in 20mM Tris buffer, pH 8.0, was applied to each membrane chromatography unit at a flow rate of 5 ml / min and the absorbance (A280) of the eluate monitored until it reached 10% of the A280 of the applied 2mg / ml BSA solution and the amount of BSA bound calculated (QB 10).

[0393] Figure 17 provides a comparison of the results. The separation device 000 with fibrous membrane functionalized with weak anion exchange (DEAE) groups (represented as Astrea DEAE in Figures 17 and 18) outperformed other commercially available devices / membranes, each of which was a traditional style chromatography membrane. The dynamic binding capacity on a volumetric basis was greater than those of the commercially available separation devices when tested. Additionally, the separation device according to this disclosure (Astrea DEAE) had by far the lowest back pressure (about 0.2 bar) compared to the commercial products.

[0394] Further comparative data is provided in Figure 18, which shows elution peaks for separation device 000 (Astrea DEAE) compared to the elution profiles for 2 of the commercially available anion-exchange membrane chromatography units: Salt tolerant AEX (run in triplicate) and Q Membrane 2. Following BSA loading to 10% breakthrough (QB10), the devices were washed with 20mM Tris buffer, pH 8.0 and bound BSA eluted with 2M NaCl at a flow rate of 5 ml / min. The 2 commercially available units tested each produced irregular / split elution peaks due to trapped air which affected liquid flow distribution within the devices. By comparison, the radial separation device(s) described herein 000 do not suffer from these deficiencies and deliver sharp and uniform elution peaks.

[0395] Example 3. A radial separation device 000 and a radial separation device 1000 were assembled with 1ml and 10 ml pleated membrane volumes respectively of a fibrous membrane functionalized with weak anion exchange (DEAE) groups. The radial separation devices were attached to a chromatography workstation equipped with a UV detector and equilibrated with 20mM Tris buffer, pH 8.0, following which 1 mg / ml BSA in 20mM Tris buffer, pH 8.0 was applied to the separation devices until the A280 of the eluates was 10% of the applied BSA solution. At this point BSA loading was halted and the separation devices flushed with 20mM Tris buffer, pH 8.0 until a steady base-line was achieved after which bound BSA was eluted with 2M NaCl. A constant flow rate of 5 MV per min was used throughout. Figure 19 shows overlayed elution peak profiles for separation device 000 and separation device 1000. The elution peak profiles are highly consistent confirming effective membrane sealing and liquid flow pathways for separation device 000 and separation device 1000. Furthermore these results demonstrate reproducible results on scale-up from a 1ml MV separation device to a 10 ml MV separation device.

[0396] Example 4. The radial separation device 000, the radial separation device 1000, and the radial separation device 2000 were assembled with 1ml, 10 ml and 100ml pleated membrane volumes respectively of a hydrophilic fibrous membrane. The separation devices were attached to a chromatography workstation equipped with a conductivity detector and flushed with water, following which 300 mM NaCl was applied to the separation devices until the conductivity of the eluates plateaued. A constant flow rate of 10 MV per min was used throughout. Figure 20 shows overlayed transition curve profiles for the separation device 000, the separation device 1000 and the separation device 2000. The conductivity profiles recorded during transition from water to 300 mM NaCl are highly consistent confirming effective membrane sealing and liquid flow pathways for the separation device 000, the separation device 1000 and the separation device 2000. Furthermore, these results demonstrate reproducible performance from a 1ml MV separation device to a 10ml MV separation device to a 100ml MV separation device.

[0397] From the foregoing, it can be seen that the present disclosure accomplishes at least all of the stated objectives.LIST OF REFERENCE CHARACTERS

[0398] The following table of reference characters and descriptors are not exhaustive, nor limiting, and include reasonable equivalents. If possible, elements identified by a reference character below and / or those elements which are near ubiquitous within the art can replace or supplement any element identified by another reference character.Table 1: List of Reference Characters

[0399] The above disclosure is not intended to refer to any single embodiment of the particular invention but encompass all possible embodiments as described in the specification and the claims. The “scope” of the present disclosure is defined by the appended claims, along with the full scope of equivalents to which such claims are entitled. The scope of the disclosure is further qualified as including any possible modification to any of the aspects and / or embodiments disclosed herein which would result in other embodiments, combinations, subcombinations, or the like that would be obvious to those skilled in the art.

Claims

CLAIMSWhat is claimed is:

1. A separation device (000) comprising: a capsule body (200); a separation media assembly (300) located at least partially within the capsule body (200); the separation media assembly (300) comprising a separation media (312) sealed within the capsule body (200), a first encapsulation member (302) and a second encapsulation member (310); wherein the separation media assembly (300) includes a center void (320); a core (600) positioned in the center void (320) of the separation media assembly (300); a lid (100); a first connector (402) proximate the lid (100) providing a first port (102) for fluid to pass through; a second connector (204) proximate the capsule body (200) providing a second port (202) for fluid to pass through; and characterised in that fluid can flow between the first connector (402) and the second connector (204) and can pass through the separation media (312) in a direction radial to a direction in which fluid passes through the second connector (204).

2. The separation device (000) according to claim 1, wherein the first encapsulation member (302) is made of an elastomeric material and is hermetically sealed to the separation media (312).

3. The separation device (000) according to any one of claims 1-2, wherein the capsule body (200) comprises a primary interface that hygienically couples the capsule body (200) with an outside region (308) of the separation media assembly (300).

4. The separation device (000) according to any one of claims 1-3, wherein the separation media (312) comprises one or more surface functionalizations.

5. The separation device (000) according to any one of claims 1-4, wherein the first connector (402) is parallel with the second connector (204).

6. The separation device (000) according to any one of claims 1-5, wherein an interior radial wall (206) of the capsule body (200) and an exterior surface of the separation media (312) provide an outer channel (212) that tapers.

7. The separation device (000) according to any one of claims 1-6, wherein an interior surface of the separation media (206) of the capsule body (200) and the core (600) provide an inner channel (214) that tapers.

8. The separation device (000) according to any one of claims 1-7, wherein the outer channel (212) and the inner channel (214) taper at a slope inverse to each other.

9. The separation device (000) according to any one of claims 1-8, wherein the interior radial wall (206) interfaces with a second encapsulation member (310) such that the flow is permitted with a controlled resistance.

10. The separation device (000) according to any one of claims 1-9, wherein the separation media (312) comprises a join (314) that is thermally bonded or encapsulated.

11. The separation device (000) according to any one of claims 1-10, wherein the capsule body (200) comprises internal ridges (216).

12. The separation device (000) according to any one of claims 1-11, wherein the core (600) maintains an equidistant flow-path reducing void volume inside the separation media (312).

13. The separation device (000) according to any one of claims 1-12, wherein the core (600) further comprises one or more fins and / or vents.

14. The separation device (000) according to any one of claims 1-13, wherein the separation media (312) comprises a hydrogel, one or more membranes, a monolith, or a matrix.

15. The separation device (000) according to claim 14, wherein the separation media (312) comprises one or more fibrous membranes or a cast membrane.

16. The separation device (000) according to any one of claims 1-15, wherein the separation media (312) is pleated or rolled.

17. The separation device (000) according to claim 15 or 16, wherein the separation media (312) comprises multiple layers, wherein the fibrous membrane comprises a primary functional layer (318A) attached to a secondary inert layer (318B).

18. The separation device (000) according to any one of claims 1-4, further comprising a conduit (2900) attached to the lid (100) and the first connector (402), wherein the first port (102) extends through the conduit (2900), wherein the first connector (402) is proximate the lid (100) via the conduit (2900), wherein the conduit (2900) redirects flow substantially 180- degrees with respect to flow that passes through the second connector (204).

19. The separation device (000) according to claim 18, wherein the lid (100) further comprises a side aperture (2126) through which the conduit (2900) passes and wherein the sealing body (2250) further comprises a conduit aperture (2290) through which the conduit passes.

20. The separation device (000) according to claim 19, wherein the first connector (402) and the second connector (204) are positioned on the same plane.

21. The separation device (000) according to any one of claims 1-20, further comprising a bleed-valve disposed on the capsule body (200).

22. The separation device (000) according to claim 20, wherein the bleed-valve is adjacent to the second connector (204).

23. The separation device (000) according to any one of claims 1-22, further comprising a first frit (800) disposed between the lid (100) and the second encapsulation member (310).

24. The separation device (000) according to any one of claims 1-23, further comprising a second frit (850) disposed between the core (600) and the capsule body (300).

25. A separation device (000) comprising: a capsule body (200) having an outlet (202) a separation media assembly (300) located at least partially within the capsule body (200); the separation media assembly (300) comprising a separation media (312) hermetically sealed within the capsule body (200) and an upper encapsulation member (302) and a lower encapsulation member (310); wherein the separation media (312) comprises one or more surface functionalizations and forms a center void 320; a core (600) positioned in the center void 320 of the separation media (312), a lid (100) having an inlet (102); characterised in that fluid that flows between the inlet and outlet and passes through the separation media (312) in a direction radial to a direction in which fluid passes through the inlet (102) and / or outlet (202).

26. A method of controlling flow through the separation device (000) of according to claim 25, comprising: flowing a fluid through the lid (100) at the inlet (102); passing the fluid through the separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which fluid passes through the inlet (102); and passing the fluid through the outlet (202).

27. A method of controlling flow through the separation device (000) of according to claim 25, comprising: flowing a fluid through the capsule body (200) at the outlet (202); passing the fluid through separation media (312);wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the inlet (102); and passing the fluid through the outlet (202).

28. A method of controlling flow through the separation device (000) of according to any one of claims 1-24, comprising: flowing a fluid through the first connector (402); passing the fluid through the separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which fluid passes through the second connector (204); and passing the fluid through the second connector (204).

29. A method of controlling flow through the separation device (000) of according to any one of claims 1-24, comprising: flowing a fluid through the capsule body (200) at the second connector (204); passing the fluid through separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the second connector (204); and passing the fluid through the first connector (402).

30. The method according to claim 29, wherein the core (600) and / or a diverter (602) direct the flow to an inner channel (214) which tapers.

31. The method according to any one of claims 29-30, wherein the fluid exits the separation media (312) into an outer channel (212) which tapers.

32. A method of purifying a target component comprising: flowing a fluid through the inlet (102) or outlet (202) of the separation device according to any of claims 1-15; passing the fluid through separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the inlet (102); separating the biomaterial from the fluid; andpassing the fluid through the outlet.

33. A method of purifying a target component comprising: flowing a fluid through the first connector (402) or second connector (204) of the separation device (000) according to any of claims 1-24; passing the fluid through separation media (312); wherein the fluid passes through the separation media (312) in a direction radial to a direction in which flow passes through the second connector (402); separating the biomaterial from the fluid; and passing the fluid through the second connector (204).

34. The method of claim 33, wherein the target component comprises an amino acid, a biomolecule or fragment thereof, a peptide, an affimer, a protein, an enzyme, a glycoprotein, an LNP, a lipopolysaccharide, an antibody and / or a fragment thereof, a nucleic acid, an organic polymer, a virus, a VLP, an EV, an exosome, a bacterium, a cell, a cell-related structure, or a mixture thereof.

35. A system for separating one or more target molecules comprising: a feed line 702, the separation device of claim 25, a discharge line 704, wherein the feed line 702 is in fluid communication with the inlet 102, and the discharge line 704 is in fluid communication with the outlet 202.

36. A system for separating one or more target molecules comprising: an upstream flow line (702), the separation device of any one of claims 1-24, a downstream flow line (704); wherein the upstream flow line (702) connects to the first connector (402) and the downstream flow line (704) connects to the second connector (204), or wherein the upstream flow line (702) connects to the second connector (204) and the downstream flow line (704) connects to the first connector (402).

37. The system of claim 36, further comprising one or more instruments and / or sensors in fluid communication with the upstream flow line (702) and / or downstream flow line (704).

38. The system of claim 37, wherein the one or more instruments and / or sensors are a chromatography workstation, a fast protein liquid chromatography system (“FPLC”) an Akta system, a process chromatography skid, a membrane chromatography skid, a high performance liquid chromatography instrument (“HPLC”), an ultra-performance liquid chromatography instrument (“UPLC”), a next generation chromatograph (“NGC”), a mass chromatograph (“MS”), a gas chromatograph (“GC”), a colorimeter, a mass spectrometer, a fluorometer, a photometer, a spectrometer, a spectrophotometer, a X-ray photoelectron spectrometer (“XPS”), a sequencing instrument, and a next generation sequencing (“NGS”) sequencing instrument, a UV detector, a visible light detector, a conductivity sensor, a pH sensor, a pressure sensor, a temperature sensor, a liquid flow sensor, a refractive index detector, a diode array detector, or any combination thereof, any or all of which may optionally be connected to a recording device.

39. The system of any one of claims 36-38, further comprising a pumping system comprising a diaphragm pump, a peristaltic pump, a syringe pump, a positive displacement pump, a quattroflow pump, a centrifugal pump and / or a lobe pump, wherein the pumping system is connected in series and in fluid communication with the separation device.

40. The system of any one of claims 36-39, further comprising a fraction collector in fluid communication with the downstream flow line (704).

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