Affinity chromatography

Abstract

The present invention relates to a method for removing chemical components from a liquid using affinity chromatography, the method comprising passing an elongated solid phase body through a conduit through which a liquid also flows.

Description

[Technical Field]

[0001] The present invention relates to a method for removing chemical components from a liquid using affinity chromatography, the method comprising passing an elongated solid phase body through a conduit through which a liquid also flows. [Background technology]

[0002] Biomolecules, such as proteins, nucleic acids, antibodies, peptides, and oligosaccharides, are highly versatile biological materials used in medicine, testing, and industrial processing. A new generation of biopharmaceuticals has yielded a variety of novel treatments for many serious diseases previously considered difficult or impossible to treat. These proteins and antibodies are typically produced in sterile fermenters using cells cultured on inexpensive nutrients such as sugars and amino acids. Alternative methods for producing proteins have also been developed using genetically modified plants and animals. While commercial production of these substances using cell cultures, plants, or animals is effective, such production has the drawback of yielding products in dilute aqueous solutions heavily contaminated with cellular by-products. This means that the actual purification of the crude material typically accounts for approximately 80% of the production cost.

[0003] The primary method for purifying proteins and antibodies uses a technique called "affinity chromatography," which uses solid materials engineered to specifically adsorb the desired substance. In this process, a cell mixture is mixed with the solid, which is then separated, washed, and finally treated with a substance that desorbs the protein from the solid, so that it can be collected in a liquid for further processing. Summary of the Invention [Problem to be solved by the invention]

[0004] Batch affinity chromatography often achieves high throughput only when performed on a large scale. Batch affinity chromatography also has long cycle times and is equipment intensive. The need for larger equipment means longer retention times, and some products may degrade during production, resulting in increased impurities and / or reduced yields. Developing and validating large-scale chromatography processes is also expensive and time-consuming.

[0005] For example, proteins are often produced by fermentation. The volume of liquid produced during the fermentation stage of protein production is large, and conventional affinity chromatography processes may require the fermentation liquid to be concentrated significantly prior to the affinity chromatography process. This can damage the desired product, and furthermore often results in a significantly higher processing flow rate in the initial loading step than in subsequent elution and / or wash steps.

[0006] Multi-column continuous synthesis methods have been developed, but these require complex valves and controls to operate. They also have the disadvantage of requiring the use of large amounts of stationary phase in parallel columns. [Means for solving the problem]

[0007] According to the present invention, there is provided a method for removing a chemical component from a liquid, comprising the steps of: a) passing an elongated object through a conduit, the conduit having a liquid inlet port and a liquid outlet port, a liquid from which the chemical component is removed passing along the conduit from the liquid inlet port to the liquid outlet port in a direction opposite to the elongated object, the conduit being configured to allow the liquid to contact the elongated object, and an affinity component attached to the elongated object, the affinity component having an affinity for the chemical component; b) washing the elongated object to remove any products present that have a lower affinity for the affinity component than the chemical component; A method is provided which includes:

[0008] The chemical component is removed from the liquid by passing the elongated object through the conduit, and the chemical component becomes associated (e.g., non-covalently bound) with the affinity component attached to the elongated object, thereby removing the chemical component from the liquid.

[0009] The elongated objects and / or liquid may be subjected to sonication (e.g. ultrasound) as they pass through the conduit. The elongated objects and / or liquid may be agitated as they pass through the conduit.

[0010] An elongated object may be passed through multiple conduits. If an elongated object is passed through multiple conduits, a liquid may be passed through multiple conduits as well. A liquid may be passed through multiple conduits in the opposite direction to the elongated object. Alternatively, a liquid may be supplied separately to each conduit. A liquid may be supplied separately to each conduit from a single liquid source. A liquid may be supplied separately to each conduit from multiple liquid sources. The liquid supplied by each of the multiple liquid sources may be different, for example, the concentration of a given reactant in the liquid may be different.

[0011] Step b) may include passing the elongated object through a washing conduit, the washing conduit having a washing liquid inlet port and a washing liquid outlet port, the washing liquid passing along the washing conduit from the washing liquid inlet port to the washing liquid outlet port in a direction opposite to the elongated object, the washing conduit being configured so that the washing liquid contacts the elongated object. Passing the elongated object through the washing conduit removes any products present on the elongated object that have a lower affinity for the affinity component than the chemical component. The chemical component remains associated (e.g., bound) with the affinity component, while any other products present in the initial liquid are washed away. The products that have a lower affinity for the affinity component than the chemical component are typically present (e.g., dissolved) in the washing liquid recovered from the washing liquid outlet port.

[0012] The elongated objects and / or cleaning fluid may be subjected to sonication (e.g., ultrasound) as they pass through the cleaning conduit. The elongated objects and / or cleaning fluid may be agitated as they pass through the cleaning conduit.

[0013] The elongated objects may be passed through multiple cleaning conduits. When the elongated objects are passed through multiple cleaning conduits, cleaning liquid may be passed through the multiple cleaning conduits as well. Cleaning liquid may be passed through the multiple cleaning conduits in the opposite direction to the elongated objects. Alternatively, cleaning liquid may be supplied separately to each cleaning conduit. Cleaning liquid may be supplied separately to each cleaning conduit from a single cleaning liquid source. Cleaning liquid may be supplied separately to each cleaning conduit from multiple cleaning liquid sources. The cleaning liquid supplied by each of the multiple cleaning liquid sources may be different, for example, the concentration of a given reactant in each cleaning liquid may be different.

[0014] The method may further comprise the step c) of recovering the chemical components from the elongated objects, which may be particularly useful when the desired product is, for example, a protein, a nucleic acid, an antibody, a peptide, a glycopeptide, a glycoprotein, or an oligosaccharide.

[0015] Step c) may include passing the elongated object through a desorption conduit, the desorption conduit having a desorption liquid inlet port and a desorption liquid outlet port, the desorption liquid passing along the desorption conduit from the desorption liquid inlet port to the desorption liquid outlet port in a direction opposite to the elongated object, the desorption conduit being configured so that the desorption liquid contacts the elongated object. Passing the elongated object through the desorption conduit desorbs the chemical component from the affinity component. The chemical component is typically present (e.g., dissolved) in the desorption liquid, which is collected from the desorption liquid outlet port.

[0016] The elongated objects and / or the desorption liquid may be subjected to sonication (e.g., ultrasound) as they pass through the desorption conduit. The elongated objects and / or the desorption liquid may be agitated as they pass through the desorption conduit.

[0017] The elongated objects may be passed through a plurality of the desorption conduits. When the elongated objects are passed through a plurality of the desorption conduits, the desorption liquid may be passed through the plurality of the desorption conduits in the same manner. The desorption liquid may be passed through the plurality of the desorption conduits in the opposite direction to the elongated objects. Alternatively, the desorption liquid may be supplied separately to each of the desorption conduits. The desorption liquid may be supplied separately to each of the desorption conduits from a single desorption liquid supply source. The desorption liquid may be supplied separately to each of the desorption conduits from a plurality of desorption liquid supplies. The desorption liquid supplied by each of the plurality of desorption liquid supplies may be different, for example, the concentration of a given reactant in each of the desorption liquids may be different.

[0018] The method may further comprise step d) recovering chemical components from the eluent recovered from the eluent outlet port. This may be achieved by extraction of the eluent recovered from the eluent outlet port. The extraction may be achieved by subjecting the eluent recovered from the eluent outlet port to chromatography. The extraction may be achieved by removing any volatile solvents present in the eluent recovered from the eluent outlet port, for example by heating and / or exposure to vacuum.

[0019] The method may further include step e) of recovering the product having a lower affinity from the wash solution, e.g., the wash solution recovered from the wash solution outlet port. This step may be particularly useful when the chemical component is, e.g., endotoxin, and the desired product is one of the products having a lower affinity for the affinity component than endotoxin. This is achieved by extracting the wash solution containing the product with a liquid having a greater affinity for the product than the product has for the wash solution. The extraction may be achieved by chromatography of the wash solution containing the product. The extraction may be achieved by removing any volatile solvents present in the wash solution from the product, e.g., by heating and / or applying a vacuum.

[0020] By using a multi-stage continuous process, each stage can be carried out in a dedicated device that optimizes the flow rates and conditions required for each stage, whereby the number of conduits can be selected such that, for example, a longer series of channels can be used in the loading stage to provide longer residence times, allowing concentration gradient effects to be fully utilized.

[0021] A plurality of said conduits, a plurality of said irrigation conduits, and a plurality of said desorption conduits may be passed through the elongated objects.

[0022] The method further includes a step f) of regenerating the affinity component. Step f) may include passing the elongated objects through a regeneration conduit, the regeneration conduit having a regenerating liquid inlet port and a regenerating liquid outlet port, the regenerating liquid passing along the regeneration conduit from the regenerating liquid inlet port to the regenerating liquid outlet port in a direction opposite to the elongated objects, the regeneration conduit configured such that the regenerating liquid contacts the elongated objects.

[0023] The elongated objects and / or the regeneration liquid may be subjected to sonication (e.g., ultrasound) as they pass through the regeneration conduit. The elongated objects and / or the regeneration liquid may be agitated as they pass through the regeneration conduit.

[0024] The elongated objects may be passed through a plurality of the regeneration conduits. When the regeneration objects are passed through a plurality of the regeneration conduits, the regeneration liquid may be passed through a plurality of the regeneration conduits as well. The regeneration liquid may be passed through a plurality of the washing conduits in the opposite direction to the elongated objects. Alternatively, the regeneration liquid may be supplied separately to each regeneration conduit. The regeneration liquid may be supplied separately to each regeneration conduit from a single regeneration liquid supply source. The regeneration liquid may be supplied separately to each regeneration conduit from a plurality of regeneration liquid supplies. The regeneration liquid supplied by each of the plurality of regeneration liquid supplies may be different, for example, the concentration of a given reactant in each regeneration liquid may be different.

[0025] Thus, during each step, the elongated objects move or can move; for example, the movement of the solid phase objects can be a movement that is considered continuous in nature (including continuous movement driven by a stepper motor that actually rotates in high-frequency steps). In some embodiments, the solid phase objects are stationary during the performance of a step and then moved to another device for performing another step. In other embodiments, the solid phase objects move intermittently during the performance of a step. The fluid phase flows during at least some of the steps, and may also flow continuously. Thereby, the present invention includes embodiments in which the solid phase objects are in contact with, e.g., surrounded by, the liquid flow during part or all of the step. The fluid may flow continuously during the step, although in some embodiments the fluid flow is discontinuous. In many embodiments, both the solid phase objects and the fluid phase move continuously from the beginning to the end of the step.

[0026] Affinity chromatography The method of the present invention can be applied in combination with known affinity chromatography techniques. Therefore, effective choices for affinity moieties, means for attaching affinity moieties to the elongated objects (or polysaccharides contained in the elongated objects), wash solutions, and means for recovering chemical components from the elongated objects are well known to those skilled in the art and are selected based on the type of chemical component. Details can be found in review articles such as "Large Scale Affinity Chromatography" by Yannis D. Clonis, Nature Biotechnology, vol. 5, December 1987, and "Affinity Chromatography: A Versatile Technique for Antibody Purification" by S. Arora, V. Saxena, and B. V. Ayyar, Methods, 116 (2017), pp. 84-94, both of which are incorporated herein in their entirety.

[0027] Chemical composition The chemical moiety may include a protein, nucleic acid, antibody, peptide, glycopeptide, polysaccharide, alkaloid, glycoprotein, or oligosaccharide. The chemical moiety may include a protein. The chemical moiety may be a protein. The chemical moiety may include an antibody. The chemical moiety may be an antibody. The chemical moiety may be an alkaloid.

[0028] Chemical constituents are typically not altered by the process, and therefore the species recovered by the methods of the present invention (e.g., as a result of the desorption step) are typically the same (i.e., have the same chemical structure) as the species present in the initial liquid.

[0029] liquid The liquid from which the chemical components are to be separated may be mammalian milk, serum, fermentation broth, ascites fluid, hybridoma, hybridoma cell lysate, plant cell lysate, mammalian cell lysate, fungal cell lysate, bacterial cell lysate, yeast cell lysate, extract of plant material, extract of fungal material, ribosomal synthesized protein.

[0030] affinity component The affinity moiety can be attached to the elongated body via a covalent bond. The affinity moiety can be attached to the elongated body via coordination and / or hydrogen bonding to a functional linker group that is itself attached to the elongated body via a covalent bond.

[0031] The affinity moiety can be attached to the elongated object using a linker group, which is typically derived from the reaction of a hydroxyl group of a polysaccharide on the elongated object with a linker agent and the resulting chemical species with the affinity moiety. Exemplary linker agents include 1,4-butanediol diglycidyl ether, cyanogen bromide, 1,1'-carbonyldiimidazole, 1,3-dibromo-2-propanol, 2,3-dibromopropanol, divinyl sulfone, epichlorohydrin, glyceraldehyde, and tresyl chloride.

[0032] The affinity component is selected based on the chemical component to be separated from the liquid: the affinity component must have a higher affinity for the chemical component than for any other product present in the liquid from which the chemical component is to be separated.

[0033] The affinity component may include a reactive dye.

[0034] The affinity component may comprise an amino acid. The affinity component includes an antibody, a peptide, a protein, a nucleic acid, a small molecule, a lectin, an antigen, or an anti-antibody.

[0035] Exemplary proteins include protein A, protein G, protein L. Exemplary lectins include concanavalin A, wheat germ agglutinin, mannan-binding protein, and jacalin.

[0036] The affinity component can include a metal ion, such as a transition metal ion. Exemplary metal ions include Co 2+ , Cu 2+ , Fe 2+ , Ni 2+ , and Zn 2+ When the affinity component comprises a metal ion, the affinity component comprises a chelating component (e.g., a solid phase particle contained within the elongate member) covalently attached to the elongate member, the chelating component chelates the metal ion. Exemplary chelating components include phenylalanine tetrazole, iminodiacetic acid, nitrilotriacetic acid, and tris(carboxymethyl)ethylenediamine.

[0037] The affinity component may include a thioether-substituted organic sulfone (eg, Thiosorb, T-gel).

[0038] The affinity component may include hydroxyapatite.

[0039] The association between the affinity component and the chemical component is typically reversible. In step a), no covalent bond is formed between the affinity component and the chemical component. The association between the affinity component and the chemical component can be by hydrogen bonding, coordinate bonding, ionic bonding, van der Waals forces, or a combination thereof.

[0040] The affinity component can be a pore of a particular size. The pore simply blocks all products larger than that particular size (the chemical component is smaller than that size). Alternatively, the pore can have a greater affinity for products having sizes within a particular range (the chemical component has a size within that range). That is, the elongated body can include a porous material with pores of a particular size. Examples include porous copolymers, such as polystyrene-divinylbenzene. The affinity component does not have to be a pore of a particular size.

[0041] Washing step The washing step b) removes any products that have a lower affinity for the affinity component than the chemical component. These products are usually impurities in the liquid from which the chemical component is separated, and which are absorbed or otherwise bound to the elongated bodies. The nature of these impurities depends on how the liquid was obtained.

[0042] Step b) may comprise contacting the elongated solid phase bodies with a wash solution. The wash solution may be water or an aqueous solution. Step b) may comprise contacting the solid phase bodies with a wash solution, the wash solution comprising a buffer. Additionally or alternatively, the wash solution may comprise a salt (e.g., NaCl or MgCl2). Additionally or alternatively, the wash solution may comprise a surfactant.

[0043] Desorption step Step c) typically involves recovering the chemical moiety from the elongated object by desorbing the chemical moiety from the affinity moiety. Step c) may comprise contacting the elongated solid phase object with a desorption solution.

[0044] The release solution may be an aqueous solution. The release solution may be a buffer solution (if a buffer solution is used in washing step b), the buffer solution in step c) will usually have a different pH than the buffer used in step b). Additionally or alternatively, the release solution may contain a salt (e.g., LiCl, NaCl, or MgCl). The release solution may contain guanidine and / or urea.

[0045] The desorption solution may be an organic solution.

[0046] The desorption solution may include an organic compound having a sufficiently high affinity for the affinity component to desorb the chemical component. The desorption solution may include imidazole, which is particularly useful when the affinity component includes metal ions.

[0047] The desorption solution may include peptides, proteins, antigens, lectins, antibodies that have a higher affinity for the affinity moiety than the chemical moiety.

[0048] elongated object The elongated object may comprise a polysaccharide, such as a cellulose or agarose material that can be cross-linked. The polysaccharide material contains hydroxyl groups to which affinity moieties can be attached, either directly or indirectly via a linker group.

[0049] The elongated object may comprise a copolymer, such as polystyrene-divinylbenzene. Such materials may be porous materials in which the affinity moieties are pores of a specific size. Such materials may have affinity components attached to them, for example, via covalent bonds.

[0050] The elongated object comprises an elongated object portion and a plurality of elongated object portions distributed along the entire length of the elongated object portion. Enclosure and Enclosure are formed from a material with a chemically inert mesh, EnclosureA plurality of solid phase particles are placed in the mesh, an affinity component is attached to the particles, and the size of the holes in the mesh and the size distribution of the solid phase particles are selected so that the particles do not pass through the mesh.

[0051] The term chemically inert is used herein to mean a polymer that is chemically unreactive and / or insoluble under the process conditions of the present method.

[0052] The mesh may be a polymer mesh, i.e., a mesh formed from a chemically inert polymer. Polymers such as polypropylene, polyethylene, polyester, polyamide (e.g., aramid), silk, etc. may be suitable. The polymer may be a fluorinated polymer or copolymer. The polymer may be polytetrafluoroethylene (PTFE) or ethylene tetrafluoroethylene polymer (ETFE). The polymer may be ETFE. The polymer may be aramid. The polymer may be polyetheretherketone (PEEK).

[0053] Alternatively, the mesh may be formed from a material selected from fiberglass, titanium, stainless steel, carbon fiber, and graphene.

[0054] The material may be formed from a mesh, for example a polymer mesh.

[0055] The pores of the mesh are typically large enough to allow unimpeded or substantially unimpeded passage of liquids and chemical components through the porous bag or tube to the particles therein. The pore size of the porous material can be less than 150 μm, less than 100 μm, less than 50 μm, or less than 25 μm. In some embodiments, the pore size is in the range of 30-80 μm, e.g., 45-65 μm.

[0056] Thin objects and Enclosure The elongated solid object may be formed from the same material. EnclosureThe two strips of material may be continuously connected to each other along both longitudinal edges of the elongated object and intermittently connected between both ends of the lateral width of the elongated object, Enclosure The two pieces of material may take the form of portions of the same piece of material folded over along the first longitudinal side of the elongate object. The two pieces of material may take the form of portions of a tubular piece of the same material formed, for example, on a circular loom.

[0057] When the material is a polymer mesh, the material is welded continuously along both longitudinal sides of the elongated object and intermittently across the width of the elongated object, Enclosure When the two strips of material are in the form of portions of the same piece of material folded over along a first longitudinal side of the strip, the material can be welded together by continuously welding along a second longitudinal side of the strip and intermittently welding across the width of the strip to form Enclosure can be formed.

[0058] Alternatively, the material may be continuously sewn, stapled, or joined along both longitudinal sides of the elongated object and continuously sewn, stapled, or joined across the width of the elongated object; Enclosure When the two strips of material are in the form of portions of the same piece of material folded over along a first longitudinal side of the elongated object, the material may be continuously sewn, stapled, or joined along a second longitudinal side of the elongated object and intermittently sewn or joined across the width of the elongated object to form Enclosure can be formed.

[0059] When two pieces of elongated material are in the form of portions of the same tubular piece of material, the material may be welded, stitched, stapled, or joined continuously across the width of the elongated object, Enclosure can be formed.

[0060] The seams or joints along the longitudinal sides or both sides of the elongated object may be adapted to provide anti-slip for the rollers, allowing the rollers to guide and / or drive the elongated material, i.e., the seams or joints along the longitudinal sides or both sides of the elongated object may comprise sprocket holes or uneven ridges or ridges.

[0061] The seams or joints may have a width in the range of 2mm to 10mm, for example in the range of 3mm to 5mm.

[0062] The solid phase particles may have a diameter of 45 to 180 μm, for example 60 to 180, or 60 to 100 μm or 150 to 180 μm. The solid phase particles may be nanoparticles, for example nanoparticles supported on an inert solid phase body having the above diameters, or nanoparticles in the form of aggregates.

[0063] The elongated object may have a length greater than 500 mm. The elongated object may have a length greater than 1000 mm. The width of the elongated object is typically in the range of 5 mm to 100 mm, for example 5 to 30 mm. The width of the elongated object may be in the range of 15 mm to 25 mm.

[0064] The elongated object may be continuous, ie, loop-shaped.

[0065] The solid phase particles are usually packed in a way that allows for free movement, which is beneficial when using ultrasound to enhance mixing of the liquid phase with the particles. Enclosure may each be only partially filled with solid phase particles. Enclosure may contain less than 90% (eg, less than 80% or less than 70%) of the maximum amount of particles that it can hold.

[0066] Enclosure is substantially the same width as the elongated object (lateral width relative to the elongated object). Enclosure Typically, there is at least one seam and / or joint extending along the longitudinal edge or edges of the elongated object, said seam or joint being EnclosureIn this case, the term "substantially" means that the width of the elongated object is Enclosure This means adding the total width of the seams and / or joints to the width of the garment.

[0067] Enclosure The length in the longitudinal direction is 5 to 50 mm (for long and thin objects, the length in the longitudinal direction is Enclosure dimensions). Enclosure The length in the longitudinal direction is 15 to 25 mm (for long and thin objects, the length in the longitudinal direction is Enclosure dimensions).

[0068] Enclosure can all be the same size. Enclosure Some groups may be smaller than others. This is useful, for example, to obtain samples of solid phase beads at various positions along the elongate member, for example, during operation. Enclosure There may be Enclosure is the other Enclosure The first group of Enclosure is the second group Enclosure It may be smaller. Enclosure Less than 20% of the total number may be in the first group. Enclosure Less than 10% of the first group may be in the first group. Enclosure may be intermittently positioned along the elongated solid object.

[0069] The solid phase particles are suitable for affinity chromatography, i.e., affinity moieties can be attached to the particles.

[0070] The solid particles may be spheres. The solid particles may be cylinders. The solid particles may be fibers. The solid beads may be irregularly shaped.

[0071] The solid phase particles may be polymer particles.

[0072] The solid phase particles may be synthetic polymers. Illustrative examples include polystyrene, polymethacrylate (e.g., Separon HEMA®), or polyacrylamide (e.g., Bio-Gel®).

[0073] The solid phase particles can be formed from natural polymers, typically polysaccharide materials that can be cross-linked, such as cellulose, dextran, or agarose. Exemplary materials include Sephadex®, Superdex®, Sepharose®, Macrosorb®, Trisacryl®, and Matrix Cellufine®.

[0074] Alternatively, the solid phase particles may be inorganic, such as a material selected from controlled pore glass, porous silica, glass fiber, and TiO2.

[0075] The affinity moiety can be attached to the solid phase particle via a covalent bond.

[0076] In a second aspect of the invention, an elongated object is provided having an affinity component attached to the elongated object, the elongated object and affinity component may have any of the features described above in relation to the first aspect of the invention.

[0077] Device The device may be a device described in WO2017 / 122009, which is incorporated herein by reference.

[0078] an affinity module comprising a conduit; a first service module operably coupled to a first side of the affinity module, the first service module supplying liquid to and / or receiving liquid from the affinity module; A system comprising: A system configured to pass a solid phase material through a conduit into an affinity module 10. The method of any preceding claim, wherein the method is performed by

[0079] The system may include two or more affinity modules arranged in series, whereby solid phase objects can pass through successive affinity modules. The system may be configured such that a liquid also passes through successive affinity modules. The system may be configured such that a liquid passes through successive affinity modules in an opposite direction to the elongated objects. Each service module supplying liquid to an affinity module may be configured to supply the liquid from the same liquid source. Two service modules each supplying liquid to an affinity module may be configured to supply the liquid from different liquid sources.

[0080] The system is a cleaning module comprising a cleaning conduit; a second service module operably coupled to the first side of the cleaning module, the second service module supplying cleaning fluid to and / or receiving cleaning fluid from the cleaning module; and The system is configured to pass the solid phase body through the washing module via the washing conduit.

[0081] The system may include two or more washing modules arranged in series, whereby the solid-phase objects can pass through the successive washing modules. The system may be configured such that a washing solution also passes through the successive washing modules. The system may be configured such that the washing solution passes through the successive washing modules in an opposite direction to the elongated objects. Each service module supplying washing solution to a washing module may be configured to supply the washing solution from the same washing solution source. Two service modules each supplying washing solution to a washing module may be configured to supply the washing solution from different washing solution sources.

[0082] The system is a desorption module comprising a desorption conduit; a third service module operably coupled to the first side of the desorption module, the third service module supplying desorption fluid to the desorption module and / or receiving desorption fluid from the desorption module; and The system is configured to pass the solid phase body through the desorption module via the desorption conduit.

[0083] The system may include two or more desorption modules arranged in series, whereby the solid phase objects can pass through the successive desorption modules. The system may be configured such that the desorption liquid also passes through the successive desorption modules. The system may be configured such that the desorption liquid passes through the successive desorption modules in a direction opposite to the elongated objects. Each service module supplying desorption liquid to a desorption module may be configured to supply the desorption liquid from the same desorption liquid source. Two service modules each supplying desorption liquid to a desorption module may be configured to supply the desorption liquid from different desorption liquid sources.

[0084] All modules (including any affinity modules, wash modules, desorption modules, and service modules) can be configured to removably couple to adjacent modules.

[0085] A first side and an opposite side of one or more affinity, washing, and / or desorption modules can each be a mating surface, and each service module can have a mating surface that can be coupled to a respective mating surface of each affinity, washing, and / or desorption module. The affinity, washing, and / or desorption modules can be removably coupled to their respective service modules.

[0086] One or more of the conduits (including the wash or desorption conduit) may comprise a solid phase inlet port and a solid phase outlet port, whereby one of the solid phase inlet port and the solid phase outlet port may be provided on a side of the affinity, wash, or desorption module, and the other of the solid phase inlet port and the solid phase outlet port may be provided on the other side of the affinity, wash, and / or desorption module.

[0087] Fluid inlet and outlet ports may be provided on the sides of the affinity, washing, and / or desorption module, thereby removably connecting and sealing to corresponding ports on adjacent modules.

[0088] The system may further comprise a retention element, wherein the affinity, wash, and / or desorption modules and their respective service modules are coupleable to the retention element.

[0089] The system may further include one or more service line modules, wherein the affinity, wash, and / or desorption modules and each service module are connectable to the service line module, which supplies and drains the service module.

[0090] The system may further comprise a solid body delivery module configured to connect in series with the first service module or affinity module, the solid body delivery module comprising a source of solid bodies.

[0091] The affinity, washing, and / or detachment module may comprise a lid portion and a body portion, the lid portion being removable from the body portion to allow access to the respective conduits, washing conduits, and / or detachment conduits.

[0092] The conduit may include a chamber, and the affinity, washing, and / or desorption module may be at least partially insertable into the chamber. protrusion whereby the chamber and protrusionand form a passage as part of the conduit. protrusion may comprise rollers over which the solid phase may pass.

[0093] The system may further comprise a drive train element for moving the solid phase body through the system.

[0094] The service module may include fluid conduits connecting with the respective fluid inlet ports of the affinity, wash, and / or desorption modules. The service module may include a fluid phase supply or discharge element, and the system may further include another service module including a power supply element. The service module may further include, or be connected to, a controller configured to control at least one of the solid phase movement rate, the fluid phase movement rate, and a process modification device responsive to one or more sensors. In this case, the sensor may include a spectrometer or other instrument capable of obtaining data collected from the fluid phase as it enters the service module.

[0095] Embodiments of the present invention are further described hereinafter with reference to the accompanying drawings. [Brief explanation of the drawings]

[0096] [Figure 1] FIG. 1 is a diagram of an exemplary apparatus that can be used to carry out the method of the present invention, and is the apparatus used to carry out Example 2. DETAILED DESCRIPTION OF THE INVENTION

[0097] As used herein, the term "removing" can mean completely removing or partially removing. Thus, it can mean removing more than 25% of a chemical component from a liquid. It can mean removing more than 75% of a chemical component from a liquid. It can mean removing more than 90% of a chemical component from a liquid. It can mean removing more than 95% of a chemical component from a liquid.

[0098] Throughout the description and claims of this specification, the terms "comprise" and "include" and variations thereof mean "including, but not limited to," and they are not intended to (and do not) exclude other parts, adjuncts, components, wholes, or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification should be understood to contemplate the plural as well as the singular, unless the context otherwise requires.

[0099] It should be understood that any feature, integer, property, compound, chemical moiety, or group described in connection with a particular aspect, embodiment, or example of the invention can also be applied to any other aspect, embodiment, or example described herein, to the extent not inconsistent therewith. All features disclosed in this specification (including any accompanying claims, abstract, and drawings), and / or all steps of any method or process similarly disclosed, can be combined in any combination, except combinations in which at least some of the features and / or steps are incompatible. The invention is not limited to the details of any of the embodiments described above. The invention includes any new or any new combination of features disclosed in this specification (including any accompanying claims, abstract, and drawings), or any new or any new combination of steps of any method or process similarly disclosed.

[0100] The reader's attention is drawn to all materials and documents related to this application that are filed contemporaneously or previously hereto and that are open to public inspection herewith, the contents of all such materials and documents being incorporated herein by reference. [Example]

[0101] Preparation of immobilized metal affinity chromatography (IMAC) ribbon 1. Prepare a saturated solution of disodium iminodiacetate in 12:5 v / v DMF / water. 2. Unwrap cotton tape (22 mm x 1 mm x 6 m) from the spool and apply it to 20 ml of 0.1% v / v surfactant in water. -1 Soak for 1 hour at 80°C. 3. Clean the tape. Using a glass, pour 20 ml of distilled water at 80°C. -1 Mix by hand for 10 minutes using the mixer. Repeat 5 times. Allow tape to dry at room temperature until dry to the touch. 4. Dry the cotton by placing it in an airtight bottle with DMF and 10% v / v 4A molecular sieves for 48 hours. 5. Immerse the DMF dried tape in anhydrous DMF at 80°C for 1 hour. A solution of POCl3 in DMF is prepared by slowly adding 24 ml of oxyphosphite (POCl3) to 6.1 liters of anhydrous DMF. 7. Heat the solution to 90C using a water bath (remove the water from the DMF solution!), add the cotton tape to the solution and continue heating at 90C for 30 minutes. 8. Remove the tape (which should be brown in color) from the DMF / POCl3 solution and wash twice with DMF. 9. Wash the tape twice with water. 10. Clean the tape with 5% w / v NaOH in water. 11. Wash the tape twice with water. 12. Clean the tape with 5% v / v acetic acid in water. 13. Wash the tape twice with water. 14. Place the chlorinated cotton tape in a flask equipped with a reflux condenser along with sufficient saturated solution of disodium iminodiacetic acid. 15. With cold water running through the condenser, heat the solution to 105-110C for 150 minutes. 16. Wash the cotton tape five times with distilled water. 17. Allow the tape to dry at room temperature for 48 hours or until dry to the touch. [Example]

[0102] Implementation of continuous chromatographic purification of lactalbumin from emulsion. Skim milk was used as a simulant of fermentation broth because it is highly consistent in composition and has a high concentration of water-soluble proteins. Fermentation broth may also require sonication (to disrupt cells and release proteins into solution) and filtration prior to affinity chromatography.

[0103] The apparatus 1 used in Example 2 is shown in Figure 1. A cotton tape 2 was passed sequentially through three conduits 3, 4, and 5. The conduits were formed by four PTFE blocks 9 shaped to provide three chambers. Three inserts 6, each with a roller 7 at its end, were inserted into each chamber. Each insert fit into a chamber to form three U-shaped conduits 3, 4, and 5. Each liquid entered each conduit 3, 4, and 5 via an inlet (not shown) located immediately below the exit point 10 of the cotton tape 2 where it entered the respective conduit. Each liquid exited each conduit 3, 4, and 5 via an inlet (not shown) located immediately below the entrance point 11 of the cotton tape 2 where it entered the respective conduit. Rollers 8 were further provided before and after the conduits to assist the tape in passing through the apparatus. 1. The functionalized cotton tape 2 is immersed in an excess of 1 M aqueous copper sulfate solution and left to stand for 30 minutes. 2. The cotton tape 2 is then removed from the solution and washed with water until the color of the wastewater disappears. 3. The tape is then passed through conduits 3, 4 and 5, the first conduit 3 accepting the flow of clarified whey, the second conduit 4 (washing conduit) accepting the flow of buffer A (pH 7 phosphate buffer, 20 mM tris(hydroxymethyl)aminomethane, 0.5 M NaCl) and the third conduit 5 (desorption conduit) accepting the flow of buffer B (pH 7 phosphate buffer, 20 mM imidazole, 0.5 M NaCl). 4. Collect the Buffer B and lactalbumin stream exiting Device 1.

Claims

1. 1. A method for removing a chemical component from an affinity liquid, comprising: a) passing an elongated object having an affinity liquid inlet port and an affinity liquid outlet port through an affinity conduit to remove a chemical component from the liquid; the liquid from which the chemical component is removed passes continuously along the affinity conduit from the affinity liquid inlet port to the affinity liquid outlet port in a direction opposite to the elongated body; the affinity conduit is configured to allow the affinity liquid to contact the elongated object; an affinity component attached to the elongated object, the affinity component having an affinity for the chemical component; b) washing the elongated object to remove any products present that have a lower affinity for the affinity component than the chemical component; the elongated object comprises an elongated object portion and a plurality of enclosures distributed along the length of the elongated object portion; the elongated object and the enclosure are made of the same material, and are formed by continuously connecting elongated material pieces along both edges in the longitudinal direction and continuously connecting both ends of their width in the lateral direction at intervals in the longitudinal direction, When the elongated material piece is composed of two pieces, the two elongated material pieces are overlapped and connected in both the longitudinal direction and the lateral direction; When the elongated piece of material is made of one piece, the elongated piece of material is folded over to connect both the longitudinal direction and the lateral direction; When the elongated material pieces are tubular, the elongated material pieces are connected in the short direction of the tubular elongated material pieces, and the enclosures are formed from a material comprising a chemically inert mesh, each enclosure contains a plurality of solid phase particles, the affinity moiety is attached to the particles, and the mesh hole size and size distribution of the solid phase particles are selected to prevent the particles from passing through the mesh; The method further includes recovering the chemical component from the elongated object, the recovering comprising passing the elongated object through a desorption conduit that desorbs the chemical component from the affinity component; the desorption conduit includes a desorption liquid inlet port and a desorption liquid outlet port, the desorption liquid passes along the desorption conduit from the desorption liquid inlet port to the desorption liquid outlet port in a direction opposite to the elongated objects, and the desorption conduit is configured so that the desorption liquid contacts the elongated objects; a release liquid is supplied to the release conduit separately from the affinity liquid supplied to the affinity conduit, the affinity liquid being different from the release liquid; A method in which the desorption conduit and the affinity conduit each comprise a chamber and a protrusion that is at least partially insertable into the chamber and has a shape corresponding to the chamber, whereby the chamber and the protrusion form a U-shaped passage as part of the conduit by inserting and fitting into the chamber.

2. The method of claim 1 , wherein the elongated object and / or the liquid are sonicated as they pass through the conduit.

3. 3. The method of claim 1, wherein step b) comprises passing the elongated object through a cleaning conduit to remove the products present on the elongated object that have a lower affinity for the affinity component than the chemical component, the cleaning conduit having a cleaning liquid inlet port and a cleaning liquid outlet port, the cleaning liquid passing along the cleaning conduit from the cleaning liquid inlet port to the cleaning liquid outlet port in a direction opposite to the elongated object, and the cleaning conduit is configured so that the cleaning liquid contacts the elongated object.

4. The method of claim 3 , wherein the elongated objects and / or the cleaning fluid are sonicated as they pass through the cleaning conduit.

5. The method of claim 1 , further comprising the step c) of recovering the chemical constituents from the elongated objects.

6. 6. The method of claim 5, wherein step c) comprises passing the elongated object through a desorption conduit that desorbs the chemical component from the affinity component, the desorption conduit having a desorption liquid inlet port and a desorption liquid outlet port, the desorption liquid passing along the desorption conduit from the desorption liquid inlet port to the desorption liquid outlet port in a direction opposite to the elongated object, and the desorption conduit is configured so that the desorption liquid contacts the elongated object.

7. 7. The method of claim 6, wherein the elongated objects and / or the desorption liquid are sonicated as they pass through the desorption conduit.

8. The method of claim 6, further comprising the step d) of recovering the chemical components from the supernatent recovered from the supernatent outlet port.

9. 4. The method of claim 3, further comprising the step e) of recovering the product having a lower affinity from the wash solution.

10. 10. The method of claim 1, wherein the elongated object is passed through a plurality of said conduits, a plurality of said cleaning conduits when dependent on claim 3, and / or a plurality of said desorption conduits when dependent on claim 6.

11. an affinity module comprising the conduit; The method of any one of claims 1 to 10, wherein the method is carried out in a system comprising: a first service module operably connected to a first side of an affinity module, the first service module supplying the liquid to the affinity module and / or receiving the liquid from the affinity module, the system being configured to pass a solid phase body through the affinity module via the conduit.

12. 12. The method of claim 1, further comprising the step f) of regenerating the affinity component.

13. 13. The method of claim 1, wherein the chemical moiety comprises a protein, a nucleic acid, an alkaloid, an antibody, a peptide, or an oligosaccharide.

14. The method of claim 1 , wherein the affinity component comprises a metal ion.

15. 12. The method of claim 11, further comprising the step c) of recovering the chemical constituents from the elongated objects, wherein step c) comprises contacting the solid phase bodies with a desorption solution, the desorption solution comprising imidazole.

16. 14. The method of any one of claims 1 to 13, wherein the affinity component comprises an antibody, a protein, a lectin, an antigen, or an anti-antibody.

17. 12. The method of claim 11, wherein step b) comprises contacting the solid phase body with a wash solution, the wash solution comprising a buffer.

18. an elongated object having an affinity component attached thereto, the elongated object comprises an elongated object portion and a plurality of enclosures distributed along the length of the elongated object portion; the elongated object and the enclosure are made of the same material, and are formed by continuously connecting elongated material pieces along both edges in the longitudinal direction and continuously connecting both ends of their width in the lateral direction at intervals in the longitudinal direction, When the elongated material piece is composed of two pieces, the two elongated material pieces are overlapped and connected in both the longitudinal direction and the lateral direction; When the elongated piece of material is made of one piece, the elongated piece of material is folded over to connect both the longitudinal direction and the lateral direction; When the elongated material pieces are tubular, the elongated material pieces are connected in the short direction of the tubular elongated material pieces, and the enclosures are formed from a material comprising a chemically inert mesh, each enclosure contains a plurality of solid phase particles, the affinity moiety is attached to the particles, the mesh hole size and the size distribution of the solid phase particles are selected to prevent the particles from passing through the mesh, and the elongated object is configured to be passed through an affinity conduit and contacted with an affinity liquid containing a chemical component; 2. An apparatus for carrying out the method of claim 1, comprising an elongated object that removes the chemical component from the affinity liquid by forming an association between the chemical component and the affinity component attached to the elongated object.

19. The device of claim 18 , wherein the affinity component comprises a metal ion.

20. 20. The device of claim 18, wherein the affinity component comprises an antibody, a peptide, a protein, a lectin, an antigen, or an anti-antibody.

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