# US 11,992,825 B2 · Adsorption medium, method for production thereof, and use thereof for purification of biomolecules > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,992,825 B2 - Document ID: US11992825B2 - Status: Granted - Kind: B2 - Application number: 15/758,012 - Filed: 2016-07-07 - Published: 2018-08-30 - Granted: 2024-05-28 - Earliest claimed: 2015-09-10 - Assignees: Sartorius Stedim Biotech GmbH - Applicants: Sartorius Stedim Biotech GmbH - Inventors: Louis Villain; Florian Taft; Jan Schwellenbach - Web page: https://patentgrep.com/patent/US11992825B2 ## CPC - B01J 20/289: bonded via a spacer - B01D 15/20: relating to the conditioning of the sorbent material - B01D 15/362: {Cation-exchange} - B01J 20/321: {consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds} - B01J 20/3219: {involving a particular spacer or linking group, e.g. for attaching an active group} - B01J 20/3278: {Polymers being grafted on the carrier} - B01J 20/328: {Polymers on the carrier being further modified} - B01J 20/3293: {Coatings on a core, the core being particle or fiber shaped, e.g. encapsulated particles, coated fibers} - C07K 1/16: by chromatography - C07K 1/18: Ion-exchange chromatography - C12N 9/2462: {Lysozyme (3.2.1.17)} - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS - 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) - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to an adsorption medium including polymeric space elements which have been bonded to the surface of the chromatography matrix, and polymer chains containing chromatographically active centers, wherein the polymer chains have been bonded to the polymeric spacer elements, as well as to a method for the production thereof, and to the use of the adsorption medium for the purification of biomolecules. ## Claims ### Claim 1 An adsorption medium comprising a chromatography matrix having a surface, the chromatography matrix selected from the group consisting of natural or synthetic fibers, polymer membranes, nonwovens and wovens; polymeric spacer elements having a molecular weight per polymeric spacer element ranging from 5000 to 2,000,000 g/mol, wherein the polymeric spacer elements are selected from the group consisting of polyamines and polyvinyl alcohols, and wherein the polymeric spacer elements are directly chemically or physically bonded to the surface of the chromatography matrix; and poly(meth)acrylate polymer chains containing chromatographically active centers, wherein the chromatographically active centers are selected from the group consisting of anionic and cationic groups and wherein the poly(meth)acrylate polymer chains are chemically bonded to the polymeric spacer elements. ### Claim 2 (depends on 1) The adsorption medium as claimed in claim 1, wherein the polymeric spacer elements do not have epoxy groups. ### Claim 3 (depends on 1) The adsorption medium as claimed in claim 1, wherein the adsorption medium has a degree of grafting ranging from 1% to 50%. ### Claim 4 (depends on 1) The adsorption medium as claimed in claim 1, wherein the chromatography matrix has a functional group density ranging from 1.5 to 30 nm−2. ### Claim 5 (depends on 1) The adsorption medium as claimed in claim 1, wherein the natural or synthetic fibers are selected from the group consisting of polyethylene terephthalate (PET), cellulose, cellulose derivatives, nylon, polyethylene (PE), polyamide (PA), polyethersulfone (PES), polyvinylidene difluoride (PVDF), polytetrafluoroethylene (PTFE), polypropylene (PP), and polysulfone. ### Claim 6 (depends on 1) The adsorption medium as claimed in claim 1, wherein the polymer membranes are selected from the group consisting of cellulose, cellulose derivatives, nylon, polyester, polyethylene (PE), polyamide (PA), polyethersulfone (PES), polyvinylidene difluoride (PVDF), polytetrafluoroethylene (PTFE), polypropylene (PP), and polysulfone. ### Claim 7 (depends on 1) The adsorption medium as claimed in claim 1, wherein the wovens are selected from the group consisting of cellulose, cellulose derivatives, nylon, polyester, polyethylene (PE), polyamide (PA), polyethersulfone (PES), polyvinylidene difluoride (PVDF), polytetrafluoroethylene (PTFE), polypropylene (PP), and polysulfone. ### Claim 8 (depends on 1) The adsorption medium as claimed in claim 1, wherein the nonwovens are selected from the group consisting of cellulose, cellulose derivatives, nylon, polyester, polyethylene (PE), polyamide (PA), polyethersulfone (PES), polyvinylidene difluoride (PVDF), polytetrafluoroethylene (PTFE), polypropylene (PP), and polysulfone. ### Claim 9 (depends on 1) The adsorption medium as claimed in claim 1, wherein the polyamine spacer elements are selected from the group consisting of polyallylarnine, polyvinylamine, polyethyleneirnine, poly(4-aminostyrene), chitosan, poly(N-methylvinylamine), poly(N-methylallylamine), poly(N,N-dimethylvinylamine), poly(N,N-dimethyliallylamine), and poly(oleylamines). ### Claim 10 (depends on 1) The adsorption medium as claimed in claim 1, wherein the poly(meth)acrylate polymer chains are selected from the group consisting of poly(1-glycerol (meth)acrylate), poly(2-glycerol (meth)acrylate), poly(hydroxy-ethyl (meth)acrylate), poly(hydroxypropyl (meth)acrylate), poly(2-aminoethyl (meth)acrylate), poly(2-aminopropyl (meth)acrylate), poly(2-(diethylamino)ethyl (meth)acrylate), polyethylene glycol (meth)acrylate), poly(hydroxybutyl (meth)acrylate), poly(glycosyloxyethyl (meth)acrylate), poly(3-(acryloyloxy)-2-hydroxypropyl (meth)acrylate), poly(3-chloro-2-hydroxypropyl (meth)acrylate), poly([tris(hydroxymethyl)] (meth)acrylate), poly(2-(4-benzoyl-3-hydroxyphenoxy)ethyl (meth)acrylates) and poly(glycidylmethacrylate). ### Claim 11 (depends on 1) The adsorption medium as claimed in claim 1, wherein the poly(meth)acrylate polymer chains are poly(glycidylmethacrylate). ### Claim 12 (depends on 1) The adsorption medium as claimed in claim 1, wherein the chromatography matrix is polyethylene terephthalate (PET) or polybutylene terephthalate (PBT) fibers, the polymeric spacer elements are poly(allylamine), poly(meth)acrylate polymer chains are poly(glycidylmethacrylate), and the chromatographically active centers are anionic groups. ### Claim 13 (depends on 1) The adsorption medium as claimed in claim 1, wherein the chromatography matrix is nylon membrane, the polymeric spacer elements are poly(allylamine), the poly(meth)acrylate polymer chains are poly(glycidylmethacrylate), and the chromatographically active centers are anionic groups. ### Claim 14 (depends on 1) The adsorption medium as claimed in claim 1, wherein the chromatography matrix is PET fibers, the polymeric spacer elements are polyvinyl alcohol, the poly(meth)acrylate polymer chains are poly(glycidylmethacrylate), and the chromatographically active centers are anionic groups. ### Claim 15 (depends on 1) The adsorption medium as claimed in claim 1, wherein the anionic groups are sulfonic acid groups. ### Claim 16 A method for producing an adsorption medium, comprising the steps: (a) providing a chromatography matrix having a surface, the chromatography matrix selected from the group consisting of natural or synthetic fibers, polymer membranes, nonwovens and wovens; (b) immobilizing polymeric spacer elements directly chemically or physically on the surface of the chromatography matrix, the polymeric spacer elements having a molecular weight per polymeric spacer element ranging from 5000 to 2,000,000 g/mol, wherein the polymeric spacer elements are selected from the group consisting of polyamines and polyvinyl alcohols; and (c) immobilizing poly(meth)acrylate polymer chains containing chromatographically active centers chemically on the polymeric spacer elements to produce the adsorption medium, wherein the chromatographically active centers are selected from the group consisting of anionic and cationic groups. ### Claim 17 (depends on 16) The method for producing an adsorption medium as claimed in claim 16, wherein step (c) comprises the steps: (c1) immobilizing a polymerization initiator on the polymeric spacer elements; (c2) carrying out a spacer element-initiated polymerization of a monomer to form immobilized poly(meth)acrylate polymer chains on the polymeric spacer elements; and (c3) optionally modifying the immobilized poly(meth)acrylate polymer chains to form chromatographically active centers on the poly(meth)acrylate polymer chains. ### Claim 18 (depends on 17) The method for producing an adsorption medium as claimed in claim 17, wherein the spacer element-initiated polymerization step (c2) is carried out by means of an atom transfer radical polymerization (ATRP). ### Claim 19 (depends on 17) The method for producing an adsorption medium as claimed in claim 17, wherein use is made in step (c2) of a polymerization solution comprising a portion of not more than 3 mol % of bifunctional monomers, based on the total amount of monomers in the solution. ### Claim 20 A method of purifying biomolecules using an adsorption medium comprising the steps of: providing an adsorption medium including a chromatography matrix having a surface, the chromatography matrix selected from the group consisting of natural or synthetic fibers, polymer membranes, nonwovens and wovens; polymeric spacer elements directly chemically or physically bound to the chromatography matrix, the polymeric spacer elements having a molecular weight per polymeric spacer element ranging from 5000 to 2,000,000 g/mol and wherein the polymeric spacer elements are selected from the group consisting of polyamines and polyvinyl alcohols; and poly(meth)acrylate polymer chains containing chromatographically active centers, wherein the chromatographically active centers are selected from the group consisting of anionic and cationic groups and wherein the poly(meth)acrylate polymer chains are chemically bound to the polymeric spacer elements; and passing a solution containing the biomolecules across the adsorption medium to obtain purified biomolecules. ### Claim 21 (depends on 20) The method as claimed in claim 20, wherein the biomolecules are proteins, peptides, amino acids, nucleic acids, viruses, virus-like particles and/or endotoxins.