# US 7,745,147 B2 · Methods and uses of antibodies in the purification of interferon > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,745,147 B2 - Document ID: US7745147B2 - Status: Granted - Kind: B2 - Application number: 11/884,324 - Filed: 2006-02-13 - Published: 2008-12-25 - Granted: 2010-06-29 - Earliest claimed: 2005-02-12 - Assignees: ViraNative AB - Inventors: Colin Robertson; Julie Smith; Erik Lundgren; Sven Lundberg - Web page: https://patentgrep.com/patent/US7745147B2 ## CPC - C07K 16/249: {Interferons} - C07K 14/56: IFN-alpha - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - 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) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract The present invention provides improved methods and uses of an immunoglobulin in the purification of an interferon composition. The methods of the invention provide for the use of a monoclonal antibody in the purification of an interferon composition comprising a plurality of interferon alpha subtypes. The use of the monoclonal antibody provides for the production of an interferon composition with a consistent proportion of interferon subtypes, providing a resulting improvement in simplicity of production of the resulting interferon composition. ## Claims ### Claim 1 A method of purifying a human multi-subtype interferon alpha composition, the method comprising the steps of: contacting crude interferon with the monoclonal antibody 3-A3-2 or a binding fragment thereof bound to a solid support, eluting the adsorbed interferon from said support, and recovering a multi-subtype interferon alpha composition comprising interferon alpha proteins of subtypes alpha 1, alpha 2, alpha 8, alpha 10, alpha 14 and alpha 21 from the eluted interferon. ### Claim 2 (depends on 1) A method as claimed in claim 1 wherein the monoclonal antibody is secreted by the hybridoma cell line having the accession number ECACC 05030201. ### Claim 3 (depends on 1) A method as claimed in claim 1 wherein the monoclonal antibody or binding fragment thereof has a heavy chain variable region amino acid sequence of SEQ ID NO: 1. ### Claim 4 (depends on 1) A method as claimed in claim 1 wherein the monoclonal antibody or binding fragment thereof has a light chain variable region amino acid sequence of SEQ ID NO:2. ### Claim 5 (depends on 1) A method as claimed in claim 1 wherein the monoclonal antibody or binding fragment thereof comprises a heavy chain amino acid sequence of SEQ ID NO:1 and a light chain variable amino acid sequence of SEQ ID NO:2. ### Claim 6 (depends on 1) A method as claimed in claim 1 wherein the solid support comprises a matrix selected from the group consisting of an agarose bead, a polystyrene bead, a silica bead, a chelating agarose bead, and a magnetic bead. ### Claim 7 (depends on 1) A method as claimed in claim 1 wherein the antibody is bound to carbohydrate, glass, silicon or another antibody. ### Claim 8 (depends on 1) A method as claimed in claim 1 wherein the antibody is bound to cyanogen bromide-activated agarose. ### Claim 9 (depends on 1) A method as claimed in claim 1 wherein the antibody is joined to the support through a linker. ### Claim 10 (depends on 1) A method as claimed in claim 1 wherein the purified multi-subtype interferon alpha composition contains interferon alpha subtype proteins at the following proportions by weight: interferon alpha 1 at 37+/−9%, alpha 2 plus alpha 21 at 30+/−7%, alpha 8 plus alpha 10 at 22+/−6%, and alpha 14 at 11+/−3%. ### Claim 11 A process for the purification of a crude human leukocyte interferon, said process comprising the steps of: (a) passing a solution of crude interferon through an immunoaffinity adsorption column, said column having monoclonal antibody 3-A3-2 or a binding fragment thereof bound to a solid support; (b) eluting antibody bound interferon alpha from the column using a buffer solution; (c) purifying the eluate by either precipitation or by ion exchange chromatography; and (d) recovering the precipitated, or ion exchange-concentrated, interferon alpha composition obtained in step (c) comprising interferon alpha proteins of subtypes alpha 1, alpha 2, alpha 8, alpha 10, alpha 14 and alpha 21. ### Claim 12 (depends on 11) The process of claim 11 wherein the monoclonal antibody or binding fragment thereof is derived from the hybridoma cell line having the accession number ECACC 05030201. ### Claim 13 (depends on 11) The process of claim 11 wherein the antibody or binding fragment thereof has a heavy chain variable region amino acid sequence of SEQ ID NO:1. ### Claim 14 (depends on 11) The process of claim 11 wherein the antibody or binding fragment thereof has a light chain variable region amino acid sequence of SEQ ID NO:2. ### Claim 15 (depends on 11) The process of claim 11 wherein the antibody or binding fragment thereof comprises a heavy chain amino acid sequence of SEQ ID NO:1 and a light chain variable amino acid sequence of SEQ ID NO:2. ### Claim 16 (depends on 11) The process of claim 11 which further comprises the step of purifying the eluate from the immunoaffinity adsorption column using an ion exchange column. ### Claim 17 (depends on 11) The process of claim 11 wherein the precipitation of step (c) comprises a further secondary precipitation step, Wherein the interferon resulting from this secondary precipitation step is further precipitated in ethanol prior to its recovery. ### Claim 18 (depends on 11) The process as claimed in claim 11 wherein the precipitate is further subject to gel filtration, with effluent fractions having absorbance at 280 nm being collected and recovered.