# US 10,308,678 B2 · Cation exchange chromatographic support and method for using same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,308,678 B2 - Document ID: US10308678B2 - Status: Granted - Kind: B2 - Application number: 15/327,161 - Filed: 2015-07-23 - Published: 2017-05-25 - Granted: 2019-06-04 - Earliest claimed: 2014-07-25 - Assignees: Asahi Kasei Medical Co., Ltd. - Applicants: Asahi Kasei Medical Co., Ltd. - Inventors: Hiroki Taniguchi; Ichiro Koguma - Web page: https://patentgrep.com/patent/US10308678B2 ## CPC - C07K 1/18: Ion-exchange chromatography - B01D 15/362: {Cation-exchange} - B01D 15/3809: {of the antigen-antibody type, e.g. protein A, G or L chromatography} - B01J 39/19: obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds - B01J 39/26: Cation exchangers for chromatographic processes - B01J 47/014: in which the adsorbent properties of the ion-exchanger are involved, e.g. recovery of proteins or other high-molecular compounds - B01J 47/12: characterised by the use of ion-exchange material in the form of ribbons, filaments, fibres or sheets, e.g. membranes (electrodialysis or electro-osmosis B01D61/42) - C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes - C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies - C08J 5/2243: {obtained by introduction of active groups capable of ion-exchange into compounds of the type C08J5/2231} - C08J 7/18: using wave energy or particle radiation - C08J 9/36: After-treatment (C08J9/22 takes precedence) - G01N 30/02: Column chromatography - G01N 30/88: Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 - G01N 30/96: using ion-exchange (G01N30/02, G01N30/90 take precedence) - C07K 2317/21: from primates, e.g. man - C08J 2323/06: Polyethene - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS - C08J: WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H (working, e.g. shaping, of plastics B29) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present invention provides a cation-exchange chromatographic support, comprising a membrane matrix and a copolymer immobilized on the surface of the membrane matrix, wherein the copolymer comprises a (meth)acrylamide-based compound and/or a (meth)acrylate-based compound as monomer units, and the support has one or more species of cation-exchange groups including at least a weak cation-exchange group at a density higher than 30 mmol/L per volume of the support. ## Claims ### Claim 1 A cation-exchange chromatographic support, comprising a membrane matrix and a copolymer immobilized on the surface of the membrane matrix, wherein the copolymer comprises a (meth)acrylamide-based compound and/or a (meth)acrylate-based compound as monomer units, and the support has one or more species of cation-exchange groups including at least a weak cation-exchange group at a density higher than 30 mmol/L per volume of the support. ### Claim 2 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein monomer units other than monomer units having the cation-exchange groups in the copolymer are neutral monomers having no charge, and the neutral monomers are a hydrophobic monomer unit and/or a hydrophilic monomer unit. ### Claim 3 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer comprises at least one species of hydrophobic monomer unit as a monomer unit. ### Claim 4 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the weak cation-exchange group is derived from any of an acrylic acid monomer, a methacrylic acid monomer, an acrylic acid compound monomer, and a methacrylic acid compound monomer. ### Claim 5 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the weak cation-exchange group is derived from a methacrylic acid monomer. ### Claim 6 (depends on 2) The cation-exchange chromatographic support according to claim 2, wherein the mass percentage of the hydrophobic monomer unit and/or the hydrophilic monomer unit in the copolymer is higher than that of the cation-exchange group-containing monomer units. ### Claim 7 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the one or more species of cation-exchange groups consist only of weak cation exchange groups. ### Claim 8 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the one or more species of cation-exchange groups are a mixture of a weak cation-exchange group and a strong cation-exchange group. ### Claim 9 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer is immobilized on the surface of the membrane matrix through covalent bond. ### Claim 10 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer comprises the (meth)acrylamide-based compound as a hydrophilic monomer unit. ### Claim 11 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer comprises the (meth)acrylate-based compound as a hydrophilic monomer unit. ### Claim 12 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer comprises 2-hydroxyethyl methacrylate as a hydrophilic monomer unit. ### Claim 13 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the membrane matrix comprises polyethylene. ### Claim 14 (depends on 13) The cation-exchange chromatographic support according to claim 13, wherein the graft ratio of the copolymer graft-polymerized onto the membrane matrix is 20 to 200%. ### Claim 15 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the copolymer substantially has no cross-linked structure. ### Claim 16 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the cation-exchange chromatographic support reduces the percentage of aggregates by 50% or more when 100 mg of antibodies including monomers and the aggregates is flow-through purified with respect to 1 mL of the support. ### Claim 17 A purification method for purifying a physiologically active substance from a mixed solution containing impurities and the physiologically active substance, the purification method comprising contacting the mixed solution with a cation-exchange chromatographic support to obtain the physiologically active substance with improved purity, the cation-exchange chromatographic support comprising a membrane matrix and a copolymer immobilized on the surface of the membrane matrix, wherein the copolymer comprises a (meth)acrylamide-based compound and/or a (meth)acrylate-based compound as monomer units, and the support has one or more species of cation-exchange groups including at least a weak cation-exchange group at a density higher than 30 mmol/L per volume of the support. ### Claim 18 (depends on 17) The purification method according to claim 17, wherein the purification method is based on a flow-through technique. ### Claim 19 (depends on 17) The purification method according to claim 17, wherein the physiologically active substance is a monomer of an antibody protein. ### Claim 20 (depends on 17) The purification method according to claim 17, wherein purification using an anion-exchange chromatographic support is performed before or after the purification step with the cation-exchange chromatographic support. ### Claim 21 (depends on 20) The purification method according to claim 20, wherein buffer replacement is not performed between the purification step with the cation-exchange chromatographic support and the purification step with the anion-exchange chromatographic support. ### Claim 22 (depends on 20) The purification method according to claim 20, wherein the hydrogen ion exponent of the buffer is changed by the addition of an acid or a base between the purification step with the cation-exchange chromatographic support and the purification step with the anion-exchange chromatographic support. ### Claim 23 (depends on 17) The purification method according to claim 17, further comprising a purification step by affinity chromatography before the purification step with the cation-exchange chromatographic support. ### Claim 24 (depends on 23) The purification method according to claim 23, wherein buffer replacement is not performed after elution of the physiologically active substance in the affinity chromatography step. ### Claim 25 (depends on 23) The purification method according to claim 23, wherein the elution buffer for use in the elution of the physiologically active substance in the affinity chromatography step is composed mainly of a monovalent acid. ### Claim 26 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the density is higher than 59 mmol/L. ### Claim 27 (depends on 17) The purification method for purifying a physiologically active substance from a mixed solution containing impurities and the physiologically active substance according to claim 17, wherein the physiologically active substance is a biomolecule. ### Claim 28 (depends on 27) The purification method for purifying a physiologically active substance from a mixed solution containing impurities and the physiologically active substance according to claim 27, wherein the biomolecule is an antibody. ### Claim 29 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the density is higher than 40 mmol/L. ### Claim 30 (depends on 1) The cation-exchange chromatographic support according to claim 1, wherein the density is higher than 49 mmol/L.