Lower chamber of expanded bed adsorption chromatoghraphy system and methods of use thereof

Abstract

Provided herein is a lower chamber of a chromatography system. The lower chamber includes a wall defining a cavity for receiving a first volume of fluid. The wall includes an upper portion having an upwardly tapered shape and a lower portion defining a collection area for collecting a plurality of chromatographic media particles. The collection area includes an annular groove. The lower chamber further includes an inlet in fluid communication with the cavity. The inlet includes an inlet opening and an inlet conduit. The inlet opening is positioned in an outer wall of the annular groove and angled to generate a vortex in the first volume of fluid when a second volume of fluid is directed into the cavity through the inlet. The plurality of chromatographic media particles are movable by the vortex toward a tapered end of the upper portion.

Description

LOWER CHAMBER OF EXPANDED BED ADSORPTION CHROMATOGHRAPHY SYSTEM AND METHODS OF USE THEREOFRELATED APPLICATION

[0001] This disclosure claims the benefit and priority of United States Provisional Patent Application Serial No. 63 / 637,515, filed April 23, 2024, which is incorporated herein by reference in its entirety.Field

[0002] The present disclosure generally relates to lower chambers of an expanded bed adsorption chromatography system and methods of use thereof.

[0003] Expanded Bed Adsorption (EBA) is a chromatographic process where desired proteins are purified from feed material. A column containing adsorbent particles is expanded by applying an upward liquid flow to the column such that a stable fluidized bed is formed with the adsorbent particles suspended in equilibrium between the particle sediment velocity and upward liquid flow velocity. Feed material is applied to the expanded bed and target proteins bind to the adsorbent particles while cell debris, cells, and contaminants pass through.

[0004] EBA generally requires a structure or system for retaining the adsorbent particles within the column. Conventional EBA systems often use a mesh screen or porous plate to retain the adsorbent particles within the column. Such mesh screens or porous plates generally include perforations in the range of 20 to 40 micrometers. The adsorbent particles are generally in the range of 50 to 150 micrometers. In an EBA system, a 40 micrometer retention mesh screen or porous plate, for example, will often clog when a fluid having particulates is applied to the column. Furthermore, flow patterns near the mesh screen or porous plate perforations often cause back mixing of the adsorbent particles.

[0005] Alternatives to a mesh screen or porous plate for retaining the adsorbent particles have been proposed. For example, a ball distributor uses a ball check valve, which prevents clogging, but produces undesirable flow distribution within the column. As another example, a large bead distributor produces sufficient flow distribution, but lacks a cleanable or hygienic design. As a further example, a moving arm distributor uses rotating arms with perforated holes, which is cleanable but generates flow patterns near the perforations thatoften cause back mixing of the adsorbent particles. An improved version of an EBA retention system is desired.

[0006] Herein provided is a lower chamber of a chromatography system. The lower chamber includes a wall defining a cavity for receiving a first volume of fluid. The wall includes an upper portion having an upwardly tapered shape and defining a first opening for receiving a rotating shaft of a rotating fluid distributor of the chromatography system. The wall further includes a lower portion defining a collection area for collecting a plurality of chromatographic media particles. The collection area includes an annular groove. The lower chamber further includes an inlet in fluid communication with the cavity. The inlet has an inlet opening and an inlet conduit. The inlet opening is positioned in an outer wall of the annular groove and angled to generate a vortex in the first volume of fluid when a second volume of fluid is directed into the cavity through the inlet. The plurality of chromatographic media particles is movable by the vortex toward a tapered end of the upper portion.

[0007] In some embodiments, the rotating shaft includes an inner channel and at least one inner channel aperture. The cavity is in fluid communication with the inner channel via the at least one inner channel aperture.

[0008] In some embodiments, the plurality of chromatographic media particles is movable from the cavity to the inner channel by the at least one inner channel aperture.

[0009] In some embodiments, the upwardly tapered shape has a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters. In some embodiments, the upwardly tapered shape includes a major diameter of about 109 millimeters, a minor diameter of about 28 millimeters, and a taper length of about 74 millimeters.

[0010] In some embodiments, the plurality of chromatographic media particles is collectable in the annular groove.

[0011] In some embodiments, the lower portion defines a second opening for receiving a mechanical seal of the rotating fluid distributor.

[0012] In some embodiments, the annular groove is positioned below the mechanical seal.

[0013] In some embodiments, the inlet conduit is positioned along a horizontal plane.

[0014] In some embodiments, the inlet is positioned in the outer wall of the annular groove at an entry angle of between about 40 degrees and about 50 degrees. In some embodiments, the inlet is positioned in the outer wall of the annular groove at an entry angle of about 45 degrees.

[0015] In some embodiments, the cavity has a volume of about 75 milliliters to about 300 milliliters.

[0016] In some embodiments, the plurality of chromatographic media particles includes resin particles.

[0017] In some embodiments, the chromatography system is an expanded bed adsorption system.

[0018] In some embodiments, the lower chamber is positioned below a chromatography column of the chromatography system.

[0019] Also provided is a chromatography system including: a baseplate of a chromatography column including a baseplate aperture extending through the surface of the baseplate; a rotating fluid distributor including a rotating shaft extending through the baseplate aperture; a bushing positioned between an inner surface of the baseplate aperture and an outer surface of the rotating shaft; and a lower chamber. The lower chamber includes a wall defining a cavity for receiving a first volume of fluid. The wall includes an upper portion having an upwardly tapered shape and defining a first opening for receiving the rotating shaft. The wall further includes a lower portion defining a collection area for collecting a plurality of chromatographic media particles. The collection area includes an annular groove. The lower chamber further includes an inlet in fluid communication with the cavity. The inlet has an inlet opening and an inlet conduit. The inlet opening is positioned in an outer wall of the annular groove and angled to generate a vortex in the first volume of fluid when a second volume of fluid is directed into the cavity through the inlet. The plurality of chromatographic media particles is movable by the vortex toward a tapered end of the upper portion.

[0020] In some embodiments, the bushing includes polytetrafluoroethylene.

[0021] In some embodiments, the rotating shaft includes an inner channel and at least one inner channel aperture. The cavity is in fluid communication with the inner channel via the at least one inner channel aperture.

[0022] In some embodiments, the plurality of chromatographic media particles is movable from the cavity to the inner channel by the at least one inner channel aperture.

[0023] In some embodiments, the upwardly tapered shape has a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters. In some embodiments, the upwardly tapered shape includes a major diameter of about 109 millimeters, a minor diameter of about 28 millimeters, and a taper length of about 74 millimeters.

[0024] In some embodiments, the plurality of chromatographic media particles is collectable in the annular groove.

[0025] In some embodiments, the lower portion defines a second opening for receiving a mechanical seal of the rotating fluid distributor.

[0026] In some embodiments, the annular groove is positioned below the mechanical seal.

[0027] In some embodiments, the inlet conduit is positioned along a horizontal plane.

[0028] In some embodiments, the inlet is positioned in the outer wall of the annular groove at an entry angle of between about 40 degrees and about 50 degrees. In some embodiments, the inlet is positioned in the outer wall of the annular groove at an entry angle of about 45 degrees.

[0029] In some embodiments, the cavity has a volume of about 75 milliliters to about 300 milliliters.

[0030] In some embodiments, the plurality of chromatographic media particles includes resin particles.

[0031] In some embodiments, the chromatography system is an expanded bed adsorption system.

[0032] In some embodiments, the lower chamber is positioned below the chromatography column.

[0033] Also provided is a method of using a lower chamber of a chromatography system for returning the plurality of chromatographic media particles from the cavity to a separate portion of the chromatography system. The method includes positioning the lower chamber below a chromatography column of the chromatography system; receiving the first volume of fluid in the cavity; directing the second volume of fluid into the cavity through the inlet thereby generating the vortex in the first volume of fluid; moving, via the vortex, theplurality of chromatographic media particles towards the tapered end of the lower chamber; and receiving the plurality of chromatographic media particles at the separate portion of the chromatography system via the at least one inner channel aperture.

[0034] In some embodiments, the second volume of fluid is directed into the cavity at a fluid rate of between about 6 and about 9.5 liters per minute.

[0035] In some embodiments, positioning the lower chamber below the chromatography column includes positioning the rotating shaft in the first opening.

[0036] In some embodiments, the first volume of fluid includes the plurality of chromatographic media particles.

[0037] In some embodiments, the method further includes collecting the plurality of chromatographic media particles in the collection area.

[0038] These and other features and advantages of the present disclosure will become apparent from the following detailed description taken together with the accompanying drawings. It should be understood, however, that the detailed description and specific examples, while indicating preferred implementations of the present disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those of skill in the art from this detailed description.Brief Description of the Drawings

[0039] For a better understanding of the various embodiments described herein, and to show more clearly how these various embodiments can be carried into effect, reference will be made, by way of example, to the accompanying drawings which show at least one example embodiment, and which are now described. The drawings are not intended to limit the scope of the teachings described herein.

[0040] FIG. 1 shows a cross-sectional perspective of a portion of a chromatography system in an exemplary embodiment of the present disclosure.

[0041] FIG. 2 shows a cross-sectional perspective view of an example lower chamber, rotating fluid distributor, and column baseplate of the chromatography system in FIG. 1.

[0042] FIG. 3A shows a cross-sectional side view of the lower chamber of FIG. 2.

[0043] FIG. 3B shows a top view of the lower chamber of FIG. 2.

[0044] FIG. 4 shows a cross-sectional side view of the example lower chamber, rotating fluid distributor, and column baseplate of FIG. 2.

[0045] FIG. 5 shows an example method of use of the lower chamber of FIG. 2.

[0046] FIG. 6 shows expansion ratios for a chromatography column in fluid communication with four different configurations of a lower chamber.

[0047] FIG. 7 shows numbers of theoretical plates for a chromatography column in fluid communication with four different configurations of a lower chamber.

[0048] FIG. 8 shows amounts of settled bed volumes required to reach a baseline volume for a chromatography column in fluid communication with four different configurations of a lower chamber.

[0049] FIG. 9 shows conductivity profiles for a chromatography column in fluid communication with four different configurations of a lower chamber.

[0050] FIG. 10 shows expansion profiles of an exemplary bushing described herein.

[0051] Further aspects and features of the example embodiments described herein will appear from the following description taken together with the accompanying drawings.Detailed Description of the Disclosure

[0052] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present application herein described for which they are suitable as would be understood by a person skilled in the art.

[0053] All features disclosed in the specification, including the claims, abstract, and drawings, and all the steps in any method or process disclosed, can be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive. Each feature disclosed in the specification, including the claims, abstract, and drawings, can be replaced by alternative features serving the same, equivalent, or similar purpose, unless expressly stated otherwise.I. Definitions

[0054] The term “chromatography system” as used herein refers to one or more components that can be used for performing chromatography. A chromatography system can include, for example, a column, a baseplate, a rotating fluid distributor, one or more volumes of fluid, chromatographic media, various tubes, channels, inlets, and outlets, and any other component known in the art. A chromatography system can further include a lower chamber as described herein. A chromatography system can be specifically configured for performingEBA chromatography, for example.

[0055] The term “volume of fluid” as used herein refers to an amount of fluid that can be present in the chromatography system. A volume of fluid can include an amount of fluid that is present in the lower chamber described herein. A volume of fluid can include an amount of fluid that is directed into the lower chamber via an inlet as described herein. A volume of fluid can include an amount of fluid that is present in a column of the chromatography system. A volume of fluid can include an amount of fluid that is present in a channel or tubing or a rotating fluid distributor of the chromatography system. A volume of fluid can include any type of fluid that is suitable for chromatographic applications known in the art. A volume of fluid can include an amount of fluid that comprises chromatographic media particles.

[0056] The term “chromatographic media particles” as used herein refers to materials used in chromatographic applications. It includes any suitable chromatographic media known in the art. Chromatographic media particles can comprise particles or beads of various materials or sizes. For example, chromatographic media particles can be in the form of resin particles. Resin-based chromatographic media particles can include, for example, tungsten carbide-agarose beads. Chromatographic media particles can be sized, for example, around 90-110 pm. Chromatographic media particles can be sized, as another example, around 110- 140 pm. Chromatographic media particles can have a characteristic density. For example, chromatographic media particles can have a density in the range of 2.8 to 3.2 g / mL. Chromatographic media particles can be configured with a specific ligand. For example, chromatographic media particles can include p-aminobenzoic acid ligands. As another example, chromatographic media particles can include diethylaminoethyl (DEAE) ligands. The chromatographic media particles can also be referred to as chromatography media particles.

[0057] The term “annular groove” as used herein refers to a groove that is generally annular in shape. An annular groove can have a width and depth suitable to accumulate chromatographic media particles. An annular groove can be generally circular, for example. An annular groove can be generally elliptical, as another example. An annular groove can be generally oval-shaped, as a further example. The center point of an annular groove can be defined generally by a rotating shaft of a rotating fluid distributor of the chromatography system.

[0058] The term “fluid communication” as used herein refers to at least two structures or portions of structures being arranged such that fluid flow between the at least two structuresor portions of structures is possible. The structures can include any structure suitable for fluid flow. For example, the structures can include a channel, a cavity, an aperture, a conduit, or portions thereof. The fluid can include any fluid suitable for chromatography systems known in the art.

[0059] The term “vortex” as used herein refers to a mass of fluid moving in a generally whirling manner. The vortex can resemble a whirlpool. The vortex can be generated in a first volume of fluid when a second volume of fluid is directed, at an angle into the annular groove described herein. The vortex is suitable for facilitating movement of particles in the vortex in a particular direction. For example, the vortex can facilitate movement of particles in a fluid from the annular groove of a lower chamber to a tapered end of a lower chamber. The fluid can include any fluid suitable for chromatography systems known in the art. The particles can include chromatographic media particles, for example.

[0060] The term “upwardly tapered shape” as used herein refers to a structure that is tapered in a generally upward direction. The structure can include an upper end with an upper diameter and a lower end with a lower diameter. The upper diameter can be smaller than the lower diameter, as to create a tapered shape between the upper and lower ends. The upwardly tapered shape can have various dimensions defining various amounts of tapering. In some embodiments, the upwardly tapered shape has a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters. In some embodiments, the ratio of the upper diameter to the lower diameter can be in the range of about 0.2 to 0.37. For example, the upwardly tapered shape can include a taper length of about 74 millimeters and a ratio of upper diameter to lower diameter of about 0.25.

[0061] The term “mechanical seal” as used herein refers to a component of a rotating fluid distributor of a chromatography system. A mechanical seal can operate to allow a rotating shaft of the rotating fluid distributor to rotate while maintaining a seal to prevent leakage of fluid from the chromatography system. A mechanical seal can generally extend through an opening defined by a lower chamber of a chromatography system as described herein. A mechanical seal can have a static face and a rotating face. It is desirable to keep chromatographic media particles away from the static face and rotating face of the mechanical seal. Accordingly, the lower chamber described herein can include a collection area positioned below the mechanical seal and configured to collect chromatographic media particles away from the static face and rotating face of the mechanical seal.

[0062] The term “entry angle” as used herein refers to the angle in a generally horizontal plane (i.e., yaw rotation) at which an inlet is positioned in an outer wall of an annular groove. For example, a circular annular groove can have an axis defined by a diameter of the annular groove, and the inlet can be positioned at a given entry angle relative to the axis defined by the diameter of the annular groove. In some embodiments, the inlet is positioned in the outer wall of the annular groove at an entry angle of between about 40 degrees and about 50 degrees. In some embodiments, the entry angle can be about 45 degrees.

[0063] The term “bushing” as used herein refers to a structure configured to line an opening. The bushing can be generally cylindrical in shape. The bushing can, for example, line an opening extending through the surface of a baseplate of the chromatography system described herein. The bushing can, for example, generally encircle a rotating shaft of a rotating fluid distributor of the chromatography system. The bushing can, in some embodiments, have a lip projecting radially outwardly at one or both ends of the bushing. The bushing can include various materials or sizes. For example, the bushing can comprise polytetrafluoroethylene.

[0064] In understanding the scope of the present disclosure, the term “comprising” and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, “including”, “having” and their derivatives. The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The term “consisting essentially of”, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0065] Further, terms of degree such as “substantially”, “about” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0066] More specifically, the term “about” means plus or minus 0.1 to 50%, 5-50%, or 10-40%, 10-20%, 10%-15%, preferably 5-10%, most preferably about 5% of the number to which reference is being made.

[0067] As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural references unless the content clearly dictates otherwise. Thus, for example, a composition containing “a compound” includes a mixture of two or more compounds.

[0068] In embodiments comprising an “additional” or “second” component, the second component as used herein is chemically different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

[0069] The term “and / or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of” or “one or more” of the listed items is used or present. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise.

[0070] The recitation of numerical ranges by endpoints herein includes all numbers and fractions subsumed within that range (e.g. 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that all numbers and fractions thereof are presumed to be modified by the term "about."II. Lower chamber, systems, and methods

[0071] In the field of Expanded Bed Adsorption (EBA) chromatography, a retention system is used for retaining chromatographic media within the chromatography column. A lower chamber of a chromatography system can accumulate chromatographic media that has been moved out of a lower end of the column due to back mixing near a rotating fluid distributor. A tapered configuration and angled inlet of the lower chamber can further allow return of the accumulated chromatographic media to the column via a vortex generated by a directed flow of fluid.

[0072] The inventors herein disclose a lower chamber of a chromatography system that accumulates chromatographic media and facilitates the return of the accumulated chromatographic media to the column.

[0073] The lower chamber’s primary components include: a wall defining a cavity, thewall having an upper portion and a lower portion; a collection area; and an inlet.

[0074] A chromatography system comprising the lower chamber may further comprise: a column and a rotating fluid distributor having a rotating shaft and at least one rotating arm.

[0075] Referring to FIG. 1 , shown therein is a cross-sectional view of a portion of an example chromatography system 100 of the present disclosure. In some embodiments, the chromatography system 100 is an EBA system. The chromatography system 100 includes a column 102, a rotating fluid distributor 108, and a lower chamber 110. FIG. 1 is provided for illustration purposes only and other configurations are possible.

[0076] The column 102 can be any suitable chromatography column appropriate for EBA chromatography known in the art. The column 102 can have an upper end 104 and a lower end 106. As shown in FIG. 1 , the rotating fluid distributor 108 can be located at the lower end 106 of the column, and the lower chamber 110 can be located below the column 102. The column 102 can further include a baseplate (not shown in FIG. 1) at the lower end 106.

[0077] Referring now to FIG. 2, shown therein is a cross-sectional perspective view of the lower chamber 110, rotating fluid distributor 108, and column baseplate 112, according to at least one embodiment.

[0078] The baseplate 112 can have an aperture 114 extending through the surface of the baseplate 112. In some embodiments, the aperture 114 extending through the surface of the baseplate is located generally in the central area of the baseplate 112.

[0079] The rotating fluid distributor 108 can be any suitable rotating fluid distributor appropriate for EBA chromatography known in the art. The rotating fluid distributor 108 can have a rotating shaft 116 having an upper portion, a middle portion, and a lower portion. One or more arms 124a-e can be coupled to the upper portion of the rotating shaft 116. Each of the arms 124a-e can define a channel (not shown in FIG. 2) that is in fluid communication with the column 102. The middle portion of the rotating shaft 116 can extend through the aperture 114 of the baseplate 112. The lower portion of the rotating shaft 116 can extend through the lower chamber 110. The rotating fluid distributor 108 can further include a mechanical seal (not shown) with a rotating face and a static face. The mechanical seal facilitates the rotational movement of the rotating shaft 116 while maintaining a seal to prevent leakage of fluid from the chromatography system 100.

[0080] The rotating shaft 116 can further include an inner channel 128 configured to allow fluid to flow through at least a portion of the rotating shaft 116. The rotating shaft 116 can further include at least one aperture 130a-b in a wall of the rotating shaft 116 (i.e., the elongated section of the rotating shaft) providing fluid communication between the inner channel 128 of the rotating shaft 116 and an environment surrounding the outside of the rotating shaft 116. In some embodiments, for example, the environment surrounding the outside of the rotating shaft 116 is the cavity 138 defined by the lower chamber 110, described in more detail below. The inner channel 128 of the rotating shaft 116 can be in fluid communication with the channel of the one or more arms 124a-e of the rotating fluid distributor 108.

[0081] The lower chamber 110 includes a wall defining a cavity, the wall having an upper portion 134 and a lower portion 136; a collection area 140; and an inlet (not shown in FIG. 2) in fluid communication with the cavity 138.

[0082] As shown in FIG. 2, the wall defines a cavity 138. In some embodiments, the cavity 138 has a volume of about 156 milliliters. However, it is understood that in some embodiments, the cavity 138 can have a volume that is greater or less than 156 milliliters. For example, in some embodiments, the cavity 138 can have a volume in the range of about 75 milliliters to about 300 milliliters. The cavity 138 is configured to receive a volume of fluid. The fluid can include any suitable fluid for chromatography systems known in the art. For example, in some embodiments, the fluid can include a buffer. In some embodiments, the fluid can include plasma and / or any suitable protein solution. In some embodiments, the fluid comprises plasma. In some embodiments, the fluid comprises plasma derivatives. In some embodiments, the fluid comprises a blood cell lysate. In some embodiments, the fluid comprises a protein solution. In some embodiments, the fluid comprises blood substitute. In some embodiments, the fluid comprises a therapeutic protein. In some embodiments, the fluid comprises haemoglobin. In some embodiments, the fluid comprises albumin. In some embodiments, the fluid comprises an immunoglobulin. In some embodiments, the fluid comprises a clotting factor. In some embodiments, the fluid comprises Factor VIII. In some embodiments, the fluid comprise fibrinogen. In some embodiments, the fluid comprises a1- antitrypsin. In some embodiments, the fluid comprises a recombinant analogue of a molecule described herein. In some embodiments, the fluid can include chromatographic media particles.

[0083] The wall can have an upper portion 134 and a lower portion 136. The upper portion 134 has an upper end 144 with an upper diameter and a lower end 146 with a lowerdiameter. In some embodiments, the upper diameter is smaller than the lower diameter such that the upper portion 134 has an upwardly tapered shape. The upper end 144 of the upper portion 134 may also be referred to as the tapered end 144. In some embodiments, the upwardly tapered shape has a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters. In some embodiments, the ratio of the upper diameter to the lower diameter can be in the range of about 0.2 to 0.37. In some embodiments, the upwardly tapered shape has a major diameter of about 109 millimeters, a minor diameter of about 28 millimeters, and a taper length of about 74 millimeters. The taper length can be measured from the bottom of the annular groove 150 to the opening 148 of the upper end 144, each of which will be described in further detail. In some embodiments, the ratio of the upper diameter to the lower diameter is about 0.25. Other suitable dimensions of the upwardly tapered shape are also possible.

[0084] The upper portion 134 defines an opening 148 at the tapered end 144 for receiving the rotating shaft 116 of the rotating fluid distributor 108. The opening 148 can be sized appropriately for the rotating shaft 116. For example, the opening 148 can be sized with sufficient clearance to allow the rotating shaft 116 to rotate freely.

[0085] The lower portion 136 defines a collection area 140 configured to collect chromatographic media particles. Referring briefly to FIG. 3A, shown therein is a cross- sectional side view of the lower chamber 110. As shown in FIG. 3A, the collection area 140 includes an annular groove 150 in which the chromatographic media particles are collected. Accordingly, the annular groove 150 is sized and shaped to collect chromatographic media particles. For example, in some embodiments, the annular groove 150 is about 10 millimeters deep and about 8 millimeters wide. In some embodiments, the inner sidewall of the annular groove 150 can be angled. For example, in some embodiments, the angle between the inner sidewall of the annular groove 150 and the bottom of the annular groove 150 can be about 110 degrees. Other suitable dimensions of the annular groove 150 are possible. In some embodiments, the annular groove 150 has a generally circular shape. In some embodiments, the annular groove 150 has a generally elliptical shape. In some embodiments, the annular groove 150 has a generally oval shape. In some embodiments, the annular groove 150 has any other suitable shape for collecting chromatographic media particles and for facilitating the generation of a vortex within the cavity 138, as described herein.

[0086] Returning now to FIG. 2, the lower portion 136 further defines an opening 152 for receiving the mechanical seal of the rotating fluid distributor 108. The opening 152 forreceiving the mechanical seal can be defined generally in the central area of the lower portion 136.

[0087] In some embodiments, the lower chamber 110 is of unitary construction. That is, the upper portion 134 and lower portion 136 are of unitary construction. In some embodiments, the upper portion 134 and lower portion 136 are separate structures that are coupled to form the lower chamber 110. In some embodiments, the upper portion 134 and lower portion 136 are removably coupled.

[0088] In some embodiments, the collection area 140 is positioned below the mechanical seal of the rotating fluid distributor 108. This positioning separates the collected chromatographic media particles from the mechanical seal and prevents abrasions at the mechanical seal surfaces. FIG. 2 is for illustration purposes only, and other configurations of the collection area 140 relative to the mechanical seal are possible. For example, a configuration that allows for the collection of chromatographic media particles away from the mechanical seal may not require the collection area 140 to be positioned below the mechanical seal.

[0089] The chromatographic media particles can be any suitable chromatographic media known in the art. In some embodiments, the chromatographic media particles are in the form of resin particles. The resin can comprise particles or beads of various materials or sizes. Examples of resins include, but are not limited to, MABDirect Resin (tungsten carbideagarose beads, median particle size distribution range: 90-110 pm; density: 2.8-3.2 g / mL; ligand: p-aminobenzoic acid) and FastLine DEAE Resin (tungsten carbide-agarose beads; median particle size distribution range: 110-140 pm; density: 2.8-3.2 g / mL; ligand: DEAE).

[0090] As shown in FIG. 2, at least a portion of the rotating shaft 116 of the rotating fluid distributor 108 extends through the cavity 138 of the lower chamber 110. In the example embodiment of FIG. 2, the lower portion of the rotating shaft 116 extends from the tapered end 144 of the upper portion 134 of the lower chamber wall through the cavity 138 towards the lower portion 136 of the lower chamber wall. The inner channel 128 of the rotating shaft 116 is in fluid communication with the cavity 138 via the at least one aperture 130a-b in the elongated section of the of the rotating shaft 116.

[0091] Returning to FIG. 3A, the lower chamber 110 includes an inlet 142 in fluid communication with the cavity 138. The inlet 142 is configured to direct fluid into the cavity 138. Accordingly, the inlet 142 can be connected to a fluid source via a channel or tube, or any other suitable connector known in the art. The inlet 142 can have an inlet opening 156and an inlet conduit 158. The inlet conduit 158 can define a longitudinal axis 160.

[0092] As shown in FIG. 3A, the inlet opening 156 is positioned in an outer wall of the annular groove 162 such that fluid that is directed into the lower chamber 110 through the inlet 142 is directed generally into the annular groove 150 of the collection area 140.

[0093] As shown in FIG. 3A, in some embodiments, the inlet conduit 158 is positioned along a horizontal plane. That is, the inlet 142 is oriented in a horizontal plane such that the longitudinal axis 160 defined by the inlet conduit 158 is generally parallel to the ground.

[0094] Referring to FIG. 3B, shown therein is a top view of the lower chamber 110. As can be seen in FIG. 3B, the inlet 142 can enter the outer wall 142 of the annular groove 150 at an angle 164 (i.e., yaw rotation in the horizontal plane). For example, in some embodiments, the inlet 142 is positioned in the outer wall 162 of the annular groove 150 at an entry angle 164 of about 45 degrees. That is, the inlet opening 156 and inlet conduit 158 can be angled into the annular groove 150 at about 45 degrees, and a certain tolerance of this angle is acceptable. In some embodiments, the inlet 142 is angled at an entry angle 164 in the range of about 40 degrees to about 50 degrees. In some embodiments, the inlet 142 is angled at an entry angle 164 in the range of about 40, about 41 degrees, about 42 degrees, about 43 degrees, about 44 degrees, about 45 degrees, about 46 degrees, about 47 degrees, about 48 degrees, about 49 degrees, or about 50 degrees.

[0095] The positioning of the inlet 142 and the angled configuration of the inlet 142 is configured to generate a vortex in the fluid in the cavity 138 when a separate volume of fluid is directed into the cavity 138 through the inlet 142. Due at least in part to the positioning of the inlet opening 156 in an outer wall 162 of the annular groove 150 and the upwardly tapered shape of the wall of the lower chamber 110, the vortex in the fluid in the cavity 138 can move the chromatographic media particles that have collected in the collection area 140 upwards in the cavity 138 towards the tapered end 144 of the upper portion 134. In some embodiments, the vortex facilitates movement of the chromatographic media particles towards the at least one aperture 130a-b in the rotating shaft 116. In some embodiments, the plurality of chromatographic media particles is moved from the cavity 138 to the inner channel 128 of the rotating shaft 116 via the at least one inner channel aperture 130a-b.

[0096] Referring to FIG. 4, shown therein is a cross-sectional side view of FIG. 2. As shown in FIG. 4, the chromatography system 100 can further include a bushing 166 positioned between an inner surface of the aperture 114 that extends through the baseplate 112 and an outer surface of the rotating shaft 116 of the rotating fluid distributor 108. The bushing 166can be sized to form a firm seal between the inner surface of the baseplate aperture 114 and the outer surface of the rotating shaft 116 to prevent chromatographic media particles from exiting the column 102. The bushing 166 can include any suitable material known in the art. For example, in some embodiments, the bushing 166 includes polytetrafluorethylene. In some embodiments, the bushing 166 comprises Teflon™. In some embodiments, the bushing 166 comprises Rulon™.

[0097] As shown in FIG. 4, each arm 124a-e (arms 124b-d not shown) of the rotating fluid distributor can have a corresponding inner channel 168a, 168e (corresponding inner channels of arms 124b-d not shown) that is in fluid communication with the inner channel 128 of the rotating shaft 116 and in fluid communication with the column 102. Accordingly, chromatographic media particles can be moved, via the vortex, from the collection area 140 towards the tapered end 144 of the cavity 138 near the one or more rotating shaft aperture 130a-b, from the tapered end 144 of the cavity 138 to the rotating shaft inner channel 128 via at least one rotating shaft aperture 130a-b, and from the rotating shaft inner channel 128 to the column 102 via at least one of the arms 124a-e of the rotating fluid distributor 108.

[0098] Referring to FIG. 5, shown therein is a flowchart diagram of an example method 500 of using the lower chamber 110 of a chromatography system 100, in accordance with at least one embodiment. In some embodiments, the method 500 is for returning the plurality of chromatographic media particles from the cavity 138 defined by the lower chamber 110 to a separate portion of the chromatography system 100. In some embodiments, the separate portion of the chromatography system 100 can include the column 102. In some embodiments, the separate portion of the chromatography system 100 can include the rotating fluid distributor 108.

[0099] Method 500 begins at 502 with positioning a lower chamber 110 below a column 102 of a chromatography system 100, in accordance with at least one embodiment described herein. The chromatography system 100 can be, for example, an EBA system. The step of positioning the lower chamber 110 below the column 102 can involve positioning a rotating shaft 116 of a rotating fluid distributor 108 of the chromatography system 100 to extend through the lower chamber 110.

[0100] At 504, operation of the chromatography system 100 begins. This step can involve receiving a first volume of fluid in the cavity 138 of the lower chamber 110. In some embodiments, the first volume of fluid can include chromatographic media particles. This step can further involve collecting chromatographic media particles in the collection area 140 ofthe lower chamber 110.

[0101] At 506, a second volume of fluid is directed into the cavity 138 through the inlet 142, thereby generating a vortex in the first volume of fluid. In some embodiments, the second volume of fluid is directed into the cavity 138 at a fluid rate of about 9.5 liters per minute. Other suitable flow rates are possible. For example, in some embodiments, the second volume of fluid is directed into the cavity 138 at a fluid rate that is less than 9.5 liters per minute. In some embodiments, the flow rate is between about 6 and about 9.5 liters per minute.

[0102] At 508, the vortex facilitates the movement of the collected chromatographic media particles in the collection area 140 upwards in the cavity 138 towards the tapered end 144 of the upper portion 134 of the lower chamber 110.

[0103] At 510, the plurality of chromatographic media particles is received at the separate portion of the chromatography system 100 via the at least one inner channel aperture 130a-b of the rotating shaft 116. This movement that is facilitated by the vortex can further facilitate the return of the chromatographic media particles to the column 102 via the at least one inner channel aperture 130a-b, the inner channel of the rotating shaft 128, and at least one of the inner channels 168a, 168e of the arms 124a-e of the rotating fluid distributor 108 of the chromatography system 100, as described herein.

[0104] Further, the definitions and embodiments described in particular sections are intended to be applicable to other embodiments herein described for which they are suitable as would be understood by a person skilled in the art. For example, in the following passages, different aspects of the disclosure are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

[0105] Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, examples of methods and materials are now described.Example 1

[0106] A lower chamber of an expanded bed adsorption (EBA) chromatography system was found to effectively collect chromatographic media particles away from the mechanical seal of a rotating fluid distributor of the EBA chromatography system and return said collected chromatographic media particles to a chromatography column of thechromatography system during operation of the chromatography system. This solution eliminates the need for a traditional chromatography retention system such as a mesh, net, perforated or porous plate, or screen.I. Introduction

[0107] A mechanical seal of a rotating fluid distributor allows the rotating fluid distributor to rotate freely under the power of a motor. It is believed that resin that enters a chamber including the mechanical seal increases abrasion and degradation of the mechanical seal. For example, resin particles can enter the mechanical seal and accelerate damage to the seal. Accordingly, a lower chamber that facilitates removal of resin from the lower chamber without damaging the mechanical seal is desired.

[0108] Four different configurations of the lower chamber were tested:1. Configuration 1 - has a collection area as described herein and an inlet at an entry angle of 45 degrees as described herein;2. Configuration 2 - has a collection area as described herein and an inlet at an entry angle of 90 degrees as described herein;3. Configuration 3 - does not have a collection area as described herein and an inlet at an entry angle of 45 degrees as described herein; and4. Configuration 4 - does not have a collection area as described herein and an inlet at an entry angle of 90 degrees as described herein.II. Procedure

[0109] In order to test the four different configurations, expansion testing, theoretical plates testing, and resin re-suspension testing were performed. Expansion testing measures proper fluidization of the resin bed and is achieved using the expansion ratio. The expansion ratio is equal to H / Ho, where H is the expanded bed height and Ho is the settled bed height. Failure to achieve an expected expansion ratio would suggest insufficient mixing at the bottom portion of the column. The acceptance criteria for the expansion testing are shown in Table 1.Table 1.

[0110] Theoretical plates testing assesses column efficiency by using a step increase in the signal. First, a low conductivity solution is injected into the column until the baseline is reached. Next, a high conductivity solution is injected until a maximum signal is reached. Next, the previous low conductivity solution is injected into the column until the baseline is reached.The number of theoretical plates is equal to t2 / o2where t equals the volume or time from the change in solution to 50% of the maximum signal and o equals half the volume or time between 15.85% and 85.15% of the maximum signal. Failure to achieve the defined success criteria indicates that the column is not performing efficiently. The acceptance criteria for the theoretical plates testing are shown in Table 2.Table 2.

[0111] Resin re-suspension testing is a qualitative test to determine whether resin is sufficiently removed from the lower chamber. Visual observation was used to provide a pass / fail rating. III. Materials and Equipment

[0112] Tables 3 to 5 detail the materials and equipment used in the study.Table 3.Table 4.Table 5.IV. Methods

[0113] Expansion T esting

[0114] 1. A settled bed height of 20 cm was used with a flow rate of 5 cm / min (2.45L / min) with a distributor RPM of 12.5.

[0115] 2. Expansion was recorded during the baseline of 0.1 M NaOH and 1 M NaOH during the measurement of the theoretical plates.

[0116] Theoretical Plates Testing

[0117] 1. A settled bed height of 20 cm was used with a flow rate of 5 cm / min (2.45L / min) with a distributor RPM of 12.5, and a 0.1 M NaOH buffer was used as a high conductivity solution and a 1 M NaOH buffer was used as a low conductivity solution.

[0118] 2. Three trials of HETP tests were conducted per lower chamber configuration.

[0119] Resin Re-suspension Testing

[0120] 1. A settled bed height of 20 cm was used with a flow rate of 5 cm / min (2.45L / min) with a distributor RPM of 12.5.

[0121] 2. Once expanded, the pump was shut off and closure of the inlet valve was delayed until the resin settled completely.

[0122] 3. Observations were made of whether the lower chamber filled with resin.

[0123] 4. The system was re-started and the effect of resin dispersion based on inlet flow through the inlet port was observed and recorded via video. A flow rate of 9.5 L / min was provided through the inlet.

[0124] 5. Steps 1-4 were repeated two additional times.

[0125] 6. Steps 1-5 were repeated for each lower chamber configuration.V. Results and Discussion

[0126] The following sections review the results of the testing to determine which lower chamber configuration has the best performance for resin re-suspension.

[0127] Expansion Testing

[0128] The expansion test uses the ratio of the expanded bed height versus the settled bed height, measured to the closest 0.5 cm, to assess the proper fluidization of the resin bed. FIG. 6 shows the expansion results for each lower chamber configuration, measured in 0.1 M NaOH. All expansions were similar, but configuration 1 (lower chamber with a collection area and an inlet angled at 45 degrees) had the best recorded expansion of 1.77. All four configurations would receive a “fair” rating according to the acceptance criteria.

[0129] Theoretical Plates Testing

[0130] To assess column efficiency, the number of theoretical plates were calculated by using a single step increase and decrease with 0.1 M NaOH and 1 M NaOH as low and high conductivity solutions, respectively. Theoretical plates measurements were completed in triplicate. FIG. 7 shows the average results for each lower chamber configuration. The highest HETP result of the tested lower chamber configurations was configuration 1 (the lower chamber with a collection area and 45 degree inlet angle), producing an HETP of 24.89. This result is nearly a “good” rating based on the HETP acceptance criteria and higher than the other three configurations, which all received a “fair” rating.

[0131] The amount of settled bed volumes (SBVs) to reach baseline for each lower chamber configuration is shown in FIG. 8. All results were similar and ranged from 3.88 to 4.3 SBV. The configuration that took the least amount of settled bed volumes was configuration 3 (the lower chamber without a collection area and an inlet positioned at 45 degrees). The configuration that took the most settled bed volumes was configuration 2 (the lower chamber with a collection area and an inlet position at 90 degrees). The lower chamber configurations without the collection area appear to have been faster overall to reach baseline compared to the lower chamber configurations with the collection area.

[0132] FIG. 9 shows the conductivity profile for each lower chamber configuration. As shown, the profiles each appear similar in slope and general shape, indicating that the results of the test are similar among the lower chamber configurations.

[0133] Resin Re-suspension Testing

[0134] The resin re-suspension test determined which lower chamber configuration performed best in preventing resin from entering the lower chamber, and which configuration performed best in facilitating the removal of the resin once it was already inside of the lower chamber. Table 6 shows the test results.Table 6.

[0135] In all configurations tested, resin was not able to enter the lower chamber when the valve was open. The bushing effectively prevented resin from entering the lower chamber (discussed in more detail in Example 2). The lower chamber with a collection area and an inlet angled at 45 degrees was the only configuration that successfully facilitated removal of the resin from the lower chamber.

[0136] Although fluid was directed into the lower chamber through the inlet at a flow rate of 9.5 L / min during testing, this does not necessarily correspond to the required flow rate to remove resin from the lower chamber.

[0137] Configurations 2 to 4 each resulted in a pile of resin on the side of the lower chamber opposite the inlet.VI. Summary

[0138] The lower chamber with configuration 1 resulted in the highest expansion of 1.77, which is a “fair” result according to the acceptance criteria. This configuration also produced the highest theoretical plates number with an HETP of 24.89, which is nearly a “good” result according to the HETP acceptance criteria. The other three configurations received a “fair” result according to the HETP acceptance criteria.

[0139] The resin re-suspension testing showed that in all four configurations, resin was not able to enter the lower chamber when the inlet flow was stopped and the inlet valve was left open, indicating that the bushing was effective in preventing backflow. When the lower chamber was purposely filled with resin, only the lower chamber with configuration 1 successfully facilitated removal of the resin. The other three configurations resulted in a pile of resin on the opposite side of the lower chamber from the inlet.Example 2

[0140] A bushing was found to prevent resin from entering the lower chamber without otherwise having an impact on the performance of the EBA chromatography system. For example, the expansion rate profile with the bushing in place was found to be comparable to previous observations. The flow rate required to clear resin from the lower chamber was also investigated.I. Introduction

[0141] A previous iteration of the bushing was sustaining abrasion and degradation effects caused by friction with the rotating distributor shaft. In some instances, the degradation was estimated to be about 0.05875 to 0.08625 mm. A bushing configuration having an increased diameter of 25.15 mm (from 25.00 mm) was investigated to reduce to potential contact and friction with the rotating fluid distributor shaft.II. Procedure

[0142] 1. The lower chamber was visually assessed for any resin accumulation while a flow of fluid was provided to the system and while no flow of fluid was provided to the system (i.e., the column was at rest).

[0143] 2. The bushing was inspected and measured to assess degradation after 24 hours of run time in the system.

[0144] 3. The expansion profile was defined in 0.1 M NaOH and compared to the expansion profile in the same column system with a previous iteration of the bushing.

[0145] 4. A flow rate for clearing the lower chamber of resin within an acceptable time frame was identified.III. Materials and Methods

[0146] This example was performed with Fastline DEAE resin. The rotating distributor was set to a rotation speed of 12.5 RPM during expansion testing.

[0147] Visual Assessment

[0148] 1. Prior to installation and testing, the bushing was visually inspected and measured using a caliper to confirm dimensions.

[0149] 2. The bushing was installed and underwent 24 hours of run time testing.

[0150] 3. The lower chamber was observed to identify the behaviour of resin inside the lower chamber throughout the testing (e.g., whether any resin entered the lower chamber after the pump had stopped, and whether the lower chamber was cleared of the resin when the pump was operating).

[0151] 4. The bushing was uninstalled and inspected and measured to assess degradation.

[0152] Expansion Testing

[0153] This testing was performed with a one pump system in 0.1 M NaOH with Fastline DEAE EBA resin.

[0154] 1. The outlet line was directed to the feed vessel such that the column recirculated the 0.1 M NaOH storage solution.

[0155] 2. The inlet valve was opened and the inlet pump was started.

[0156] 3. After 4 SBV passed through the column, stability of the bed expansion was confirmed and bed height was recorded.

[0157] 4. If the bed expansion was still increasing after 4 SBV, the procedure continued until the bed height was stable. The number of SBV passed was estimated by the time elapsed, as shown in Table 7.Table 7.

[0158] 5. The inlet pump flow rate was increased to a flow rate of 2.45 L / min (e.g., equivalent to a velocity of 5 cm / min), then steps 3-4 were repeated.

[0159] 6. Step 5 was repeated for 6 cm / min, 7 cm / min, 8 cm / min, 9 cm / min, and 10 cm / min and any additional desired flow rates.

[0160] 7. The system was shut down, the inlet pump was reduced back to a reasonable flow rate for regular operation, and column storage was performed, as required.

[0161] Flow Rate Verification

[0162] 1. Resin was introduced into the column and the lower chamber was visually inspected for any resin. If the chamber was clear of resin, resin was introduced into the lower chamber by temporarily running the pump in reverse at a low flow rate.

[0163] 2. The pump was started at the first test flow rate and a stopwatch was used to record the time it took to clear the chamber at the selected flow rate.

[0164] 3. If it took more than 30 seconds to clear the lower chamber, the pump was stopped and moved to the next flow rate.

[0165] 4. The pump was allowed to run for an extra 5 minutes and it was observed whether the resin moved back into the lower chamber.

[0166] 5. Steps 1 to 4 were repeated for all flow rates to be tested.IV. Results and Discussion

[0167] Visual Inspection

[0168] The average diameter of the bushing was 25.13 mm before installation and 25.13 mm after 24 hours of operation. Evidence of resin moving between the bushing and the shaft was observed by black markings left on the inner diameter of the bushing. However, no resin was observed in the lower chamber during normal operation.

[0169] Expansion Testing

[0170] The expansion test used the ratio of the expanded bed height versus settled bed height, each measured to the closest 0.5 cm to assess the proper fluidization of the resin bed. The expansion ratio study was conducted in a column with a settled bed height of 18.0 cm. FIG. 10 shows the expansion results measured in 0.1 M NaOH at various flow rates. As shown, the expansion profile observed with the bushing is comparable to previous expansion results observed in the same column with a previous iteration of the bushing.

[0171] Flowrate Verification

[0172] The flowrate verification test determined the flowrate required to clear resin from the lower chamber.

[0173] Resin was not introduced to the lower chamber during setup nor during normal column operation. Accordingly, resin was forced into the lower chamber by temporarily running the pump in a reverse flow direction. Volumetric flowrates of 2.1 L / min to 4.2 L / min were found to be ineffective in removing the resin from the lower chamber within 30 seconds. The volumetric flowrate was increased to 7.5 L / min with the distributor running at 12.5 RPM to completely clear the lower chamber of resin in under 30 seconds.

[0174] At this flowrate, there is a risk of losing resin from the column due to overexpansion depending on the buffer or solution being used. Accordingly, the flowrate should be reduced as needed and sufficient time should be provided to clear the resin from the lower chamber. Alternatively, the lower chamber can be cleared by providing high flowrate pulses to the lower chamber.V. Summary

[0175] The bushing with a diameter of 25.13 mm did not impact the standard performance of the EBA column during operation. The bushing effectively prevented any resin from entering the lower chamber during operation. If resin is introduced in the lower chamber, a suitable flowrate should be selected based on the solution used to provide a sufficient flowrate to clear the lower chamber of resin without losing resin from the column due to overexpansion.

[0176] While the present disclosure has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

[0177] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present disclosure is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

Claims

Claims:1 . A lower chamber of a chromatography system, the lower chamber comprising: a wall defining a cavity for receiving a first volume of fluid, the wall comprising: an upper portion having an upwardly tapered shape and defining a first opening for receiving a rotating shaft of a rotating fluid distributor of the chromatography system; and a lower portion defining a collection area for collecting a plurality of chromatographic media particles, the collection area comprising an annular groove; and an inlet in fluid communication with the cavity, the inlet having an inlet opening and an inlet conduit, the inlet opening positioned in an outer wall of the annular groove and angled to generate a vortex in the first volume of fluid when a second volume of fluid is directed into the cavity through the inlet, wherein the plurality of chromatographic media particles is movable by the vortex toward a tapered end of the upper portion.

2. The lower chamber of claim 1 , wherein the rotating shaft comprises an inner channel and at least one inner channel aperture, the cavity being in fluid communication with the inner channel via the at least one inner channel aperture.

3. The lower chamber of claim 2, wherein the plurality of chromatographic media particles is movable from the cavity to the inner channel by the at least one inner channel aperture.

4. The lower chamber of any one of claims 1 to 3, wherein the upwardly tapered shape comprises a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters.

5. The lower chamber of any one of claims 1 to 4, wherein the plurality of chromatographic media particles is collectable in the annular groove.

6. The lower chamber of any one of claims 1 to 5, wherein the lower portion defines a second opening for receiving a mechanical seal of the rotating fluid distributor.

7. The lower chamber of claim 6, wherein the annular groove is positioned below the mechanical seal.

8. The lower chamber of any one of claims 1 to 7, wherein the inlet conduit is positioned along a horizontal plane.

9. The lower chamber of any one of claims 1 to 8, wherein the inlet is positioned in the outer wall of the annular groove at an entry angle of about between about 40 degrees and about 50 degrees.

10. The lower chamber of any one of claims 1 to 9, wherein the cavity has a volume of about 75 milliliters to about 300 milliliters.

11. The lower chamber of any one of claims 1 to 10, wherein the plurality of chromatographic media particles comprise resin particles.

12. The lower chamber of any one of claims 1 to 11 , wherein the chromatography system is an expanded bed adsorption system.

13. The lower chamber of any one of claims 1 to 12, wherein the lower chamber is positioned below a chromatography column of the chromatography system.

14. A chromatography system comprising: a baseplate of a chromatography column comprising a baseplate aperture extending through the surface of the baseplate; a rotating fluid distributor comprising a rotating shaft extending through the baseplate aperture; a bushing positioned between an inner surface of the baseplate aperture and an outer surface of the rotating shaft; and a lower chamber comprising:a wall defining a cavity for receiving a first volume of fluid, the wall comprising: an upper portion having an upwardly tapered shape and defining a first opening for receiving the rotating shaft; and a lower portion defining a collection area for collecting a plurality of chromatographic media particles, the collection area comprising an annular groove; and an inlet in fluid communication with the cavity, the inlet having an inlet opening and an inlet conduit, the inlet opening positioned in an outer wall of the annular groove and angled to generate a vortex in the first volume of fluid when a second volume of fluid is directed into the cavity through the inlet, wherein the plurality of chromatographic media particles is movable by the vortex toward a tapered end of the upper portion.

15. The chromatography system of claim 14, wherein the bushing comprises polytetrafluoroethylene.

16. The chromatography system of claim 14 or 15, wherein the rotating shaft comprises an inner channel and at least one inner channel aperture, the cavity being in fluid communication with the inner channel via the at least one inner channel aperture.

17. The chromatography system of any one of claims 14 to 16, wherein the plurality of chromatographic media particles is movable from the cavity to the inner channel by the at least one inner channel aperture.

18. The chromatography system of any one of claims 14 to 17, wherein the upwardly tapered shape comprises a major diameter of between about 90 and about 120 millimeters, a minor diameter of between about 25 and about 32 millimeters, and a taper length of between about 69 and about 93 millimeters.

19. The chromatography system of any one of claims 14 to 18, wherein the plurality of chromatographic media particles is collectable in the annular groove.

20. The chromatography system of any one of claims 14 to 19, wherein the lower portion defines a second opening for receiving a mechanical seal of the rotating fluid distributor.

21. The chromatography system of claim 20, wherein the annular groove is positioned below the mechanical seal.

22. The chromatography system of any one of claims 14 to 21 , wherein the inlet conduit is positioned along a horizontal plane.

23. The chromatography system of any one of claims 14 to 22, wherein the inlet is positioned in the outer wall of the annular groove at an entry angle of between about 40 degrees and between about 50 degrees.

24. The chromatography system of any one of claims 14 to 23, wherein the cavity has a volume of about 75 milliliters to about 300 milliliters.

25. The chromatography system of any one of claims 14 to 24, wherein the plurality of chromatographic media particles comprise resin particles.

26. The chromatography system of any one of claims 14 to 25, wherein the chromatography system is an expanded bed adsorption system.

27. The chromatography system of any one of claims 14 to 26, wherein the lower chamber is positioned below the chromatography column.

28. A method of using a lower chamber of a chromatography system according to any one of claims 1 to 12 for returning the plurality of chromatographic media particles from the cavity to a separate portion of the chromatography system, the method comprising: positioning the lower chamber below a chromatography column of the chromatography system; receiving the first volume of fluid in the cavity; directing the second volume of fluid into the cavity through the inlet thereby generating the vortex in the first volume of fluid;moving, via the vortex, the plurality of chromatographic media particles towards the tapered end of the lower chamber; and receiving the plurality of chromatographic media particles at the separate portion of the chromatography system via the at least one inner channel aperture.

29. The method of claim 28, wherein the second volume of fluid is directed into the cavity at a fluid rate of between about 6 and about 9.5 liters per minute.

30. The method of claim 28 or 29, wherein positioning the lower chamber below the chromatography column comprises positioning the rotating shaft in the first opening.31 . The method of any one of claims 28 to 30, wherein the first volume of fluid comprises the plurality of chromatographic media particles.

32. The method of any one of claims 28 to 31 , further comprising collecting the plurality of chromatographic media particles in the collection area.

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