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 extracting chemical components from a liquid using affinity chromatography. The method includes passing an elongated solid phase through a conduit through which a liquid also flows.

Background Art

[0002] Biomolecules such as proteins, nucleic acids, antibodies, peptides, and oligosaccharides are highly versatile biological substances and are used in medicine, testing, and industrial processing. New generation biopharmaceuticals have created various novel treatment methods for many serious diseases that were previously considered difficult or impossible to treat. These proteins and antibodies are usually produced in a sterile fermenter using cells cultured with inexpensive nutrients such as sugars and amino acids. Another method for producing proteins has also been developed by using genetically modified plants and animals. The actual production of these substances using cell culture, plants, or animals is effective, but the production has the drawback that the product is obtained in a dilute aqueous solution with a large amount of cell by-products mixed in. This means that the actual purification of the crude raw material usually exceeds about 80% of the production cost.

[0003] The main method for purifying proteins and antibodies uses a technique called "affinity chromatography". This uses a solid material that is engineered to specifically adsorb the desired substance. In this process, the cell mixture is mixed with the solid, then the solid is separated, washed, and finally treated with a substance that desorbs the protein from the solid, so that the protein can be collected in a liquid for further processing.

Summary of the Invention

Problems to be Solved by the Invention

[0004] Batch affinity chromatography often achieves high yields only when implemented on a large scale. Batch affinity chromatography also involves long cycle times and is equipment-intensive. The need for larger equipment means longer retention times, which can lead to degradation of some products during production, increased impurity, and / or reduced yield. Deploying and validating large-scale chromatography processes is also costly 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 in conventional affinity chromatography processes, it may be necessary to concentrate the fermentate considerably before the affinity chromatography process. This can damage the desired product and, furthermore, often results in a significantly larger processing flow rate in the initial filling stage than in the subsequent elution and / or washing stages.

[0006] While multi-column sequential synthesis methods have been developed, these methods require the operation of complex valves and controls. 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, a method for extracting chemical components from a liquid, a) A conduit is provided to pass an elongated object through, the conduit having a liquid inlet port and a liquid outlet port, and a liquid from which a chemical component is extracted flows along the conduit from the liquid inlet port to the liquid outlet port in the opposite direction to the elongated object, the conduit is configured such that the liquid comes into contact with the elongated object, an affinity component is attached to the elongated object, and the affinity component has affinity for the chemical component, b) Washing an elongated object to remove a product that has a lower affinity for the affinity component than the chemical component. A method including this is provided.

[0008] Chemical components are extracted from a liquid by passing a slender object through a conduit. The chemical components associate (for example, by non-covalent bonds) with affinity components attached to the slender object, thereby extracting the chemical components from the liquid.

[0009] Elongated objects and / or liquids may be subjected to sonic treatment (e.g., ultrasound) as they pass through the conduit. Elongated objects and / or liquids may be agitated as they pass through the conduit.

[0010] An elongated object can be passed through multiple conduits. When an elongated object is passed through multiple conduits, a liquid can similarly be passed through multiple conduits. The liquid can be passed through multiple conduits in the opposite direction to the elongated object. Alternatively, the liquid can be supplied to each conduit separately. The liquid can be supplied to each conduit separately from a single liquid supply source. The liquid supplied by each of the multiple liquid supply 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 comprising a washing fluid inlet port and a washing fluid outlet port, wherein the washing fluid flows along the washing conduit from the washing fluid inlet port to the washing fluid outlet port in the opposite direction from the elongated object, and the washing conduit may be configured so that the washing fluid comes into contact with the elongated object. Passing the elongated object through the washing conduit removes products present on the elongated object that have a lower affinity to the affinity component than the chemical component. The chemical component remains associated (e.g., bound) with the affinity component, and any other products that were present in the initial liquid are washed away. Products with a lower affinity to the affinity component than the chemical component are usually present (e.g., dissolved) in the washing fluid recovered from the washing fluid outlet port.

[0012] Elongated objects and / or cleaning fluids may be subjected to ultrasonic treatment (e.g., ultrasound) as they pass through the cleaning conduit. Elongated objects and / or cleaning fluids may be agitated as they pass through the cleaning conduit.

[0013] An elongated object can be passed through multiple cleaning conduits. When an elongated object is passed through multiple cleaning conduits, the cleaning solution can similarly be passed through multiple cleaning conduits. The cleaning solution can be passed through multiple cleaning conduits in the opposite direction to the elongated object. Alternatively, the cleaning solution can be supplied to each cleaning conduit separately. The cleaning solution can be supplied to each cleaning conduit separately from a single cleaning solution source. The cleaning solution supplied by each of the multiple cleaning solution sources may be different, for example, the concentration of a given reactant in each cleaning solution may be different.

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

[0015] Step c) may include passing an elongated object through a desorption conduit, the desorption conduit comprising a desorption inlet port and a desorption outlet port, wherein the desorption fluid flows along the desorption conduit from the desorption inlet port to the desorption outlet port in the opposite direction to the elongated object, and the desorption conduit is configured such that the desorption fluid comes into contact with the elongated object. By passing the elongated object through the desorption conduit, the chemical components are desorbed from the affinity components. The chemical components are typically present (e.g., dissolved) in the desorption fluid recovered from the desorption outlet port.

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

[0017] An elongated object can be passed through multiple desorption conduits. When an elongated object is passed through multiple desorption conduits, the desorption liquid can similarly be passed through multiple desorption conduits. The desorption liquid can be passed through multiple desorption conduits in the opposite direction to the elongated object. Alternatively, the desorption liquid can be supplied separately to each desorption conduit. The desorption liquid can be supplied separately to each desorption conduit from a single desorption liquid supply source. The desorption liquid supplied by each of the multiple desorption liquid supply sources may be different, for example, the concentration of a given reactant in each desorption liquid may be different.

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

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

[0020] By employing a multi-stage continuous process, each stage can be carried out using dedicated equipment optimized for the flow rate and conditions required for each stage. This allows the number of conduits to be selected such that, for example, a longer series of channels can be used in the filling stage to provide a longer residence time, thereby fully utilizing the concentration gradient effect.

[0021] A slender object can be passed through the multiple conduits, the multiple cleaning conduits, and the multiple detachment conduits.

[0022] The method further comprises step f) regenerating affinity components. Step f) may include passing an elongated object through a regeneration conduit, the regeneration conduit comprising a regeneration fluid inlet port and a regeneration fluid outlet port, wherein the regeneration fluid flows along the regeneration conduit from the regeneration fluid inlet port to the regeneration fluid outlet port in the opposite direction to the elongated object, and the regeneration conduit is configured such that the regeneration fluid comes into contact with the elongated object.

[0023] Elongated objects and / or regenerating fluid may be subjected to ultrasonic treatment (e.g., ultrasound) as they pass through the regenerating conduit. Elongated objects and / or regenerating fluid may be agitated as they pass through the regenerating conduit.

[0024] An elongated object can be passed through multiple regeneration conduits. When a regenerated object is passed through multiple regeneration conduits, the regeneration liquid can similarly be passed through multiple regeneration conduits. The regeneration liquid can be passed through multiple cleaning conduits in the opposite direction to the elongated object. Alternatively, the regeneration liquid can be supplied to each regeneration conduit separately. The regeneration liquid can be supplied to each regeneration conduit separately from a single regeneration liquid supply source. The regeneration liquid supplied by each of the multiple regeneration liquid supply sources may be different, for example, the concentration of a given reagent in each regeneration liquid may be different.

[0025] Thus, during each step, the elongate object can move or be movable, for example, the movement of the solid-phase object can be a movement that is considered to be practically continuous (including in fact a continuous movement driven by a stepper motor rotating in high-frequency steps). In some embodiments, the solid-phase object remains stationary during the performance of the step and is then moved to another device for performing another step. In another embodiment, the solid-phase object moves intermittently during the performance of the step. The fluid phase can flow during at least a part of the steps and can also flow continuously. Thereby, the invention includes embodiments in which the solid-phase object is in contact with the liquid flow, for example surrounded by the flow, during a part or the whole of the step. The fluid can flow continuously during the step, but in some embodiments, the fluid flow is discontinuous. In many embodiments, both the solid-phase object and the fluid phase move continuously from the start 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. Thus, effective options for the affinity moiety, the means for attaching the affinity moiety to the elongate object (or the polysaccharide contained in the elongate object), the washing solution, and the means for recovering the chemical components from the elongate object are well known to those skilled in the art and are selected based on the type of chemical components. Details can be found in review articles such as Nature Biotechnology, vol 5, December 1987 - Large Scale Affinity Chromatography by Yannis D Clonis, and Methods, 116(2017), 84-94 - Affinity Chromatography: A versatile technique for antibody purification by S Arora, V Saxena and B V Ayyar, both of which are incorporated herein in their entirety.

[0027] Chemical components Chemical components may include proteins, nucleic acids, antibodies, peptides, glycopeptides, polysaccharides, alkaloids, glycoproteins, or oligosaccharides. Chemical components may include proteins. Chemical components may be proteins. Chemical components may include antibodies. Chemical components may be antibodies. Chemical components may be alkaloids.

[0028] The chemical composition is generally not altered by this process. Therefore, the chemical species recovered by the method of the present invention (for example, as a result of the elimination step) are usually the same as the chemical species present in the initial liquid (i.e., they have the same chemical structure).

[0029] liquid The liquids from which chemical components will be separated may include mammalian milk, serum, fermentation fluid, ascites, hybridoma, hybridoma cell lysate, plant cell lysate, mammalian cell lysate, fungal cell lysate, bacterial cell lysate, yeast cell lysate, plant substance extract, fungal substance extract, and ribosomal synthesis proteins.

[0030] affinity component Affinity components can be attached to elongated objects via covalent bonds. Affinity components can be attached to elongated objects via coordination bonds and / or hydrogen bonds to functional linker groups to which they themselves are attached via covalent bonds.

[0031] Affinity components can be attached to elongated objects using linker groups. Linker groups are typically derived from the reaction of hydroxyl groups of polysaccharides on the elongated object with a linker agent, and the resulting chemical species with affinity components. Exemplary linker agents include 1,4-butanediol diglycidyl ether, cyanide bromide, 1,1'-carbonyldiimidazole, 1,3-dibromo-2-propanol, 2,3-dibromopropanol, divinyl sulfone, epichlorohydrin, glyceraldehyde, and tresilchloride.

[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 products present in the liquid from which it is being separated.

[0033] The affinity components may include reactive dyes.

[0034] Affinity components may include amino acids. Affinity components may include antibodies, peptides, proteins, nucleic acids, small molecules, lectins, antigens, or anti-antibodies.

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

[0036] The affinity components may include metal ions, such as transition metal ions. Exemplary metal ions include Co 2+ Cu 2+ Fe 2+ Ni 2+ , and Zn 2+ This includes: If the affinity component contains a metal ion, the affinity component includes a chelating component (for example, solid-phase particles contained within the elongated member) attached to the elongated member by covalent bonds. The chelating component chelates the metal ion. Exemplary chelating components include phenylalanine tetrazole, iminodiacetic acid, nitrilotriacetic acid, and tris(carboxymethyl)ethylenediamine.

[0037] The affinity components may include thioether-substituted organic sulfones (e.g., thiosorb, T-gel).

[0038] The affinity component may include hydroxyapatite.

[0039] The association between affinity components and chemical components is usually reversible. In step a), no covalent bond is formed between the affinity component and the chemical component. The association between affinity components and chemical components can be a hydrogen bond, a coordination bond, an ionic bond, a van der Waals force, or a combination thereof.

[0040] The affinity component may be a pore of a specific size. The pore simply does not allow all products larger than that specific size to enter (the chemical component is smaller than that size). Alternatively, the pore may have a greater affinity for products within a specific size range (the chemical component is within that size range). That is, an elongated object may contain a porous material having pores of a specific size. Examples include porous copolymers, such as polystyrene-divinylbenzene. The affinity component does not necessarily have to be a pore of a specific size.

[0041] Cleaning step Washing step b) removes any products that have a lower affinity for the affinity component than the chemical component. These products are typically impurities in the liquid from which the chemical component is separated, and these impurities are absorbed into elongated objects or separately linked. The nature of these impurities depends on how the liquid was obtained.

[0042] Step b) may include contacting the elongated solid phase with a cleaning solution. The cleaning solution may be water or an aqueous solution. Step b) may include contacting the solid phase with a cleaning solution, wherein the cleaning solution contains a buffer. In addition to or instead of a buffer, the cleaning solution may contain a salt (e.g., NaCl or MgCl2). In addition to or instead of a buffer, the cleaning solution may contain a surfactant.

[0043] Detachment step In step c), the chemical components are typically recovered from the elongated object by desorbing them from the affinity portion. Step c) may include bringing the elongated solid phase into contact with the desorption solution.

[0044] The desorption solution may be an aqueous solution. The desorption solution may also be a buffer solution (if a buffer solution is used in washing step b), the buffer solution in step c) is usually at a different pH than the buffer used in step b). In addition to or instead of the above, the desorption solution may contain a salt (e.g., LiCl, NaCl, or MgCl2). The desorption solution may contain guanidine and / or urea.

[0045] The aforementioned elimination solution may also be an organic solution.

[0046] The elimination solution may contain an organic compound having a sufficiently high affinity for the affinity component to eliminate the chemical component. The elimination solution may contain imidazole, which is particularly effective when the affinity component contains metal ions.

[0047] The elimination solution may contain peptides, proteins, antigens, lectins, and antibodies having a higher affinity for the affinity moiety than the chemical components.

[0048] elongated object The elongated object may contain polysaccharides, such as crosslinkable cellulose or agarose substances. The polysaccharide substance contains hydroxyl groups to which affinity components can be attached either directly or indirectly via linker groups.

[0049] The elongated objects may contain copolymers, 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] An elongated object may comprise an elongated portion and a plurality of surrounding portions distributed along the entire length of the elongated portion, wherein the surrounding portions are formed from a material having a chemically inert mesh, a plurality of solid-phase particles are placed in each surrounding portion, affinity components are attached to the particles, and the size of the mesh holes 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 does not react and / or is insoluble under the process conditions of this method.

[0052] The mesh may be a polymer mesh, i.e., a mesh formed from a chemically inert polymer. Suitable polymers may include polypropylene, polyethylene, polyester, polyamide (e.g., aramid), and silk. The polymer may also be a fluorinated polymer or copolymer. The polymer may also be polytetrafluoroethylene (PTFE) or ethylenetetrafluoroethylene polymer (ETFE). The polymer may also be ETFE. The polymer may also be aramid. The polymer may also be polyetheretherketone (PEEK).

[0053] Alternatively, the mesh can be formed from a material selected from glass fiber, titanium, stainless steel, carbon fiber, or graphene.

[0054] The material may be formed from a mesh, such as a polymer mesh.

[0055] The pores of the mesh are typically large enough to allow liquids and chemical components to pass through the porous bag or tube to the internal particles without obstruction or substantially obstruction. The pore size of the porous material may 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 to 80 μm, for example, in the range of 45 to 65 μm.

[0056] The elongated object and the surrounding portion can be formed from the same material. The elongated solid-phase object may comprise two elongated material pieces integrally connected to form an elongated portion and a plurality of surrounding portions. The two material pieces can be continuously connected to each other along both longitudinal edges of the elongated object, and intermittently, continuously connected between the two ends of the width of the elongated object to form the surrounding portion. The two material pieces may take the form of parts of the same material piece folded along the first longitudinal side of the elongated object. The two material pieces may take the form of parts of the same tubular material piece formed, for example, on a circular loom.

[0057] If the material is a polymer mesh, the material can be welded continuously along both longitudinal sides of an elongated object, and intermittently, continuously between the two ends of the width of the elongated object to form a surrounding portion. If two pieces of the elongated material take the form of a part of the same material folded along the first longitudinal side of the elongated object, the material can be welded continuously along the second longitudinal side of the elongated object, and intermittently, continuously between the two ends of the width of the elongated object to form a surrounding portion.

[0058] Alternatively, a surrounding portion can be formed by continuously stitching, stapling, or joining materials along both longitudinal sides of an elongated object, or by continuously stitching, stapling, or joining materials between the two ends of the width of the elongated object. If two pieces of an elongated material take the form of parts of the same material folded along the first longitudinal side of the elongated object, the material can be continuously stitched, stapling, or joining materials along the second longitudinal side of the elongated object, or intermittently stitching or joining materials between the two ends of the width of the elongated object to form a surrounding portion.

[0059] When two elongated pieces of material take the form of a part of the same tubular piece of material, the material can be welded, sutured, stapled, or joined continuously between the two ends of the width of the elongated object to form a surrounding portion.

[0060] A seam or joint along the longitudinal side or both sides of an elongated object may be suitable for preventing roller slippage, allowing the roller to guide and / or drive the elongated object. That is, the seam or joint along the longitudinal side or both sides of an elongated object may comprise sprocket holes or uneven ridges or protrusions.

[0061] The seams or joints may have a width ranging from 2 mm to 10 mm, for example, from 3 mm to 5 mm.

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

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

[0064] The elongated object may be continuous, that is, it may be loop-shaped (ring-shaped).

[0065] Solid phase particles are typically packed in a way that allows for free movement. This is beneficial when using ultrasound to enhance mixing of the liquid phase and particles. Therefore, each surrounding area may be filled only partially with solid phase particles. The surrounding area may accommodate less than 90% (e.g., less than 80% or less than 70%) of the maximum amount of particles it can hold.

[0066] The surrounding portion may have substantially the same width as the elongated object (the lateral dimension of the surrounding portion relative to the elongated object). Typically, there is at least one seam and / or joint extending along the longitudinal edge or both edges of the elongated object, and the seam or joint forms the surrounding portion. In this context, the term "substantially" means that the width of the elongated object is the sum of the width of the surrounding portion and the width of the seam and / or joint.

[0067] The surrounding portion may have a longitudinal length of 5 to 50 mm (the longitudinal dimension of the surrounding portion relative to an elongated object). The surrounding portion may have a longitudinal length of 15 to 25 mm (the longitudinal dimension of the surrounding portion relative to an elongated object).

[0068] The surrounding portions may all be the same size, or some may be smaller than others. This is useful, for example, for obtaining samples of solid-phase beads at various positions along an elongated member, for example, during operation. Two groups of surrounding portions may exist, and the surrounding portions of each group may be the same size as the other surrounding portions of that group. The surrounding portions of the first group may be smaller than those of the second group. Less than 20% of the total number of surrounding portions may belong to the first group. Less than 10% of the surrounding portions may belong to the first group. The surrounding portions of the first group may be intermittently arranged along an elongated solid-phase object.

[0069] Solid-phase particles are suitable for affinity chromatography; that is, affinity components can be attached to the particles.

[0070] The solid phase particles may be spherical. The solid phase particles may be cylindrical. The solid phase particles may be fibers. The solid phase beads may have an irregular shape.

[0071] The solid-phase particles may also 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] Solid-phase particles can be formed from natural polymers, typically crosslinkable polysaccharides, 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 materials selected from, for example, pore-controlled glass, porous silica, glass fibers, and TiO2.

[0075] Affinity components can be attached to solid-phase particles via covalent bonds.

[0076] In a second aspect of the present invention, an elongated object is provided to which an affinity component is attached. The elongated object and the affinity component may have any of the above-described features relating to the first aspect of the present invention.

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

[0078] A compatibility module equipped with a conduit, A first service module operably connected to a first side of an affinity module, the first service module supplying liquid to and / or receiving liquid from the affinity module A system that is equipped with, A system configured to pass a solid phase through a conduit to an affinity module. The method according to any of the claims, as implemented in [location].

[0079] The system may comprise two or more affinity modules arranged in a sequence, thereby allowing a solid phase to pass through the sequence of affinity modules. The system may also be configured so that a liquid passes through a sequence of affinity modules. The system may be configured so that a liquid passes through a sequence of affinity modules in the opposite direction to that of an elongated object. 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 equipped with a cleaning conduit, A second service module operably connected to a first side of a cleaning module, the second service module supplying cleaning fluid to the cleaning module and / or receiving cleaning fluid from the cleaning module. It may further be equipped with, The system is configured to pass a solid phase through a cleaning conduit to the cleaning module.

[0081] The system may comprise two or more consecutively arranged cleaning modules, thereby allowing the solid phase to pass through the consecutive cleaning modules. The system may also be configured so that the cleaning fluid passes through the consecutive cleaning modules in the opposite direction to the elongated object. Each service module supplying cleaning fluid to a cleaning module may be configured to supply the cleaning fluid from the same cleaning fluid source. Two service modules, each supplying cleaning fluid to a cleaning module, may be configured to supply the cleaning fluid from different cleaning fluid sources.

[0082] The system is A deactivation module equipped with a deactivation conduit, A third service module operably connected to a first side of a desorption module, the third service module supplying desorption fluid to the desorption module and / or receiving desorption fluid from the desorption module. It may further be equipped with, The system is configured to pass a solid phase through a desorption conduit to a desorption module.

[0083] The system may comprise two or more desorption modules arranged in a sequence, thereby allowing the solid phase to pass through the sequence of desorption modules. The system may also be configured so that the desorption fluid passes through the sequence of desorption modules in the opposite direction to that of the elongated object. Each service module supplying desorption fluid to a desorption module may be configured to supply the desorption fluid from the same desorption fluid source. Two service modules, each supplying desorption fluid to a desorption module, may be configured to supply the desorption fluid from different desorption fluid sources.

[0084] All modules (including all affinity modules, cleaning modules, deactivation modules, and service modules) can be configured to be detachably connected to adjacent modules.

[0085] One or more affinity, cleaning, and / or desorption modules have a first side and the opposite side that can be a mating surface, and each service module has a mating surface that can be connected to the respective mating surface of each affinity, cleaning, and / or desorption module. The affinity, cleaning, and / or desorption modules can be removably connected to each service module.

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

[0087] Fluid inlet and outlet ports may be provided on the sides of affinity, cleaning, and / or deactivation modules, thereby being detachably connected to and sealed to corresponding ports on adjacent modules.

[0088] The system may further include retention elements, affinity, cleaning, and / or detachment modules, and their respective service modules, which can be connected to the retention elements.

[0089] The system may further comprise one or more service conduit modules, affinity, cleaning, and / or deactivation modules, and each service module may be connectable to the service conduit modules, which may supply and discharge to the service modules.

[0090] The system may further comprise a solid phase delivery module configured to be connected in series with a first service module or affinity module, the solid phase delivery module comprising a solid phase source.

[0091] The affinity, cleaning, and / or deactivation module may comprise a lid portion and a body portion, the lid portion being removable from the body portion, thereby providing access to the respective conduits, cleaning conduits, and / or deactivation conduits.

[0092] The conduit may comprise a chamber, and an affinity, cleaning, and / or desorption module may comprise a projection that is at least partially insertable into the chamber, thereby the chamber and the projection forming a passage as part of the conduit. The projection may comprise rollers over which a solid phase can pass.

[0093] The system may further include drive-train elements for moving a solid phase through the system.

[0094] A service module may comprise fluid conduits connecting to the fluid inlet ports of each of the affinity, cleaning, and / or desorption modules. A service module may comprise a fluid phase supply or discharge element, and the system may further comprise yet another service module comprising a power supply element. A service module may further comprise, or be connected to, a control device, which is configured to control the moving speed of a solid phase, the moving speed of a fluid phase, and at least one of a process modifier that responds to one or more sensors. In this case, the sensors include a spectrometer or other instrument that can obtain data collected from the fluid phase as it enters the service module.

[0095] Embodiments of the present invention will be described further below with reference to the accompanying drawings. [Brief explanation of the drawing]

[0096] [Figure 1] This is an exemplary apparatus that can be used to carry out the method of the present invention, and is a diagram of the apparatus used to carry out Example 2. [Modes for carrying out the invention]

[0097] As used herein, the term “remove” may mean to remove completely or to remove partially. Therefore, it may mean removing more than 25% of the chemical components from the liquid. It may mean removing more than 75% of the chemical components from the liquid. It may mean removing more than 90% of the chemical components from the liquid. It may mean removing more than 95% of the chemical components from the liquid.

[0098] Throughout this description and claims, the terms “equipped with” and “included,” and their variations, mean “included, but not limited to,” and they are not intended (and do not exclude) other parts, appendices, components, completes, or steps. Throughout this description and claims, the singular includes the plural unless otherwise required by context. In particular, where the indefinite article is used, the specification should be understood to intend both the singular and the plural unless otherwise required by context.

[0099] Features, wholes, properties, compounds, chemical parts, or groups described in connection with a particular aspect, embodiment, or example of the present invention should be understood to be applicable to any other aspect, embodiment, or example described herein, insofar as they do not conflict therewith. All features disclosed herein (including all appended claims, abstract, and drawings) and / or all steps of any similarly disclosed methods or processes can be combined in any combination, except for combinations in which at least some of those features and / or steps are incompatible. The present invention is not limited to any details of any embodiment described above. The present invention encompasses any new features or any new combinations of features disclosed herein (including all appended claims, abstract, and drawings), and any new steps or any new combinations of any similarly disclosed methods or processes.

[0100] The reader's attention is directed to all materials and documents filed concurrently with or prior to this specification in connection with this application and made available for public viewing together with this specification, the contents of all such materials and documents are incorporated herein by reference. [Examples]

[0101] Preparation of immobilized metal affinity chromatography (IMAC) ribbons Prepare a saturated solution of disodium iminodiacetate in a 1.12:5v / v DMF / water mixture. 2. Unwind the cotton tape (22mm x 1mm x 6m) from the spool and add 20mlg of 0.1% v / v surfactant in water. -1 Soak in 80°C for 1 hour. 3. Clean the tape. Using a glass, rinse with 20 ml of 80°C distilled water. -1 Mix by hand for 10 minutes using [the product name]. Repeat 5 times. Let the tape dry at room temperature until it feels dry. 4. Dry the cotton by placing it in an airtight bottle with DMF and 10% v / v 4A molecular sieves for 48 hours. 5. Soak the dried DMF tape in anhydrous DMF at 5.80°C for 1 hour. A DMF solution of POCl3 is prepared by slowly adding 24 ml of oxyphosphate salt (POCl3) to 6.1 liters of anhydrous DMF. 7. Heat the solution to 90°C using a water bath (to remove water from the DMF solution!), add cotton tape to the solution, and continue heating at 90°C for 30 minutes. 8. Remove the tape (which should be brown) from the DMF / POCl3 solution and wash it twice with DMF. 9. Wash the tape twice with water. 10. Wash the tape with a 5% w / v NaOH aqueous solution. 11. Wash the tape twice with water. 12. Wash the tape with a 5% v / v acetic acid solution. 13. Wash the tape twice with water. 14. Place the chlorinated cotton tape, along with a sufficient saturated solution of disodium imino2acetate, into a flask equipped with a reflux condenser. 15. Heat the solution to 105-110°C for 150 minutes while supplying cold water to the condenser. 16. Wash the cotton tape five times with distilled water. 17. Allow the tape to dry at room temperature for 48 hours or until it feels dry. [Examples]

[0102] Performing serial chromatography purification of lactalbumin from emulsion Since the emulsion has a very similar composition and a high concentration of water-soluble proteins, skimmed emulsion was used as a simulated substance for the fermentation liquid. The fermentation liquid may also require sonication (to disrupt cells and release proteins into solution) and filtration before affinity chromatography.

[0103] The apparatus 1 used in Example 2 is shown in Figure 1. Cotton tape 2 was sequentially passed through three conduits 3, 4, and 5. The conduits were formed by four PTFE blocks 9 shaped to have three chambers. Three inserts 6, each having a roller 7 at one end, were inserted into each chamber. Each insert fits into the chamber to form three U-shaped conduits 3, 4, and 5. Each liquid flows into each conduit 3, 4, and 5 via an inlet (not shown) just below the outlet 10 of the cotton tape 2 entering each conduit. Each liquid flows out of each conduit 3, 4, and 5 via an inlet (not shown) just below the inlet 11 of the cotton tape 2 entering each conduit. Rollers 8 were further provided at the front and back of the conduits to assist the tape in passing through the apparatus. 1. Immerse the functionalized cotton tape 2 in an excess 1M copper sulfate aqueous solution and let it stand for 30 minutes. 2. Next, remove the cotton tape 2 from the solution and wash it with water until the wastewater is no longer colored. 3. Next, pass the tape through conduits 3, 4, and 5. The first conduit 3 allows the flow of clarified whey, the second conduit 4 (washing conduit) allows 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) allows the flow of buffer B (pH 7 phosphate buffer, 20 mM imidazole, 0.5 M NaCl). 4. Collect the flow of buffer B and lactalbumin coming out of device 1.

Claims

1. 1. A method for removing a chemical component from a liquid, comprising: a) passing an elongated object through a conduit to remove a chemical component from a liquid, the conduit having a liquid inlet port and a liquid outlet port, the 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 such that the liquid contacts 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 comprising:

2. 10. 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 washing conduit to remove the products present on the elongated object that have a lower affinity for the affinity component than the chemical component, 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, and the washing conduit is configured so that the washing liquid contacts the elongated object.

4. 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. 5. The method according to 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. 7. The method of claim 5 or 6, further comprising the step d) of recovering the chemical components from the supernatent recovered from the supernatent outlet port.

9. 9. The method of claim 1, further comprising the step e) of recovering the product with lower affinity from the wash solution.

10. 10. The method of claim 1, wherein the elongated object is passed through a plurality of the conduits, a plurality of the cleaning conduits, and / or a plurality of the desorption conduits.

11. 11. The method of claim 1, wherein the elongated object comprises an elongated object portion and a plurality of surrounding portions distributed along the length of the elongated object portion, the surrounding portions being formed from a material comprising a chemically inert mesh, each surrounding portion containing a plurality of solid phase particles, the affinity component being attached to the particles, and the size of the holes in the mesh and the size distribution of the solid phase particles being selected so that the particles do not pass through the mesh.

12. an affinity module comprising the conduit; a first service module operably coupled to a first side of the affinity module, the first service module supplying the liquid to the affinity module and / or receiving the liquid from the affinity module; A system comprising: a system configured to pass a solid phase body through the conduit and into the affinity module; 12. The method according to any one of claims 1 to 11, wherein the method is carried out in

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

14. 14. The method of any one of claims 1 to 13, wherein the chemical moiety comprises a protein, a nucleic acid, an alkaloid, an antibody, a peptide, or an oligosaccharide.

15. 15. The method of any one of claims 1 to 14, wherein the affinity moiety comprises a metal ion.

16. 16. The method of claim 15, further comprising the step c) of recovering the chemical moiety from the elongated object, wherein step c) comprises contacting the solid phase object with a wash solution, the wash solution comprising imidazole.

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

18. 18. The method of claim 1, wherein step b) comprises contacting the solid phase body with a wash solution, the wash solution comprising a buffer.

19. An elongated object to which an affinity component is attached.

20. 20. The elongated object of claim 19, wherein the elongated object comprises an elongated object portion and a plurality of surrounding portions distributed along the entire length of the elongated object portion, the surrounding portions being formed from a material comprising a chemically inert mesh, each surrounding portion containing a plurality of solid phase particles, the affinity component being attached to the particles, and the size of the holes in the mesh and the size distribution of the solid phase particles being selected so that the particles do not pass through the mesh.

21. 21. The elongated object of claim 19 or 20, wherein the affinity moiety comprises a metal ion.

22. 21. The elongated object according to claim 19 or 20, wherein the affinity moiety comprises an antibody, a peptide, a protein, a lectin, an antigen, or an anti-antibody.

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