# US 6,020,182 A · Subunit respiratory syncytial virus vaccine preparation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 6,020,182 A - Document ID: US6020182A - Status: Granted - Kind: A - Application number: 08/679,060 - Filed: 1996-07-12 - Granted: 2000-02-01 - Earliest claimed: 1996-07-12 - Assignees: Connaught Laboratories Limited - Inventors: George A. Cates; Sonia E. Sanhueza; Raymond P. Oomen; Michel H. Klein - Web page: https://patentgrep.com/patent/US6020182A ## CPC - C07K 14/005: from viruses - A61K 39/12: Viral antigens - A61K 39/155: Paramyxoviridae, e.g. parainfluenza virus - A61P 11/04: for throat disorders - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 31/12: Antivirals - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - G01N 33/56983: {Viruses} - A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) - A61K 2039/55505: {Inorganic adjuvants} - A61K 2039/55511: {Organic adjuvants} - A61K 2039/55577: {Saponins; Quil A; QS21; ISCOMS} - C07K 2319/00: Fusion polypeptide - C12N 2760/18521: Viruses as such, e.g. new isolates, mutants or their genomic sequences - C12N 2760/18522: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2760/18534: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/18551: Methods of production or purification of viral material - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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 fusion (F) protein, attachment (G) protein and matrix (M) protein of respiratory syncytial virus (RSV) are isolated and purified from respiratory syncytial virus by mild detergent extraction of the proteins from concentrated virus, loading the protein onto a hydroxyapatite or other ion-exchange matrix column and eluting the protein using mild salt treatment. The F, G and M proteins, formulated as immunogenic compositions, are safe and highly immunogenic and protect relevant animal models against respiratory syncytial virus. ## Claims ### Claim 1 A mixture of purified fusion protein (F), attachment protein (G) and matrix (M) protein of respiratory syncytial virus in the relative proportions of: F from about 40 to about 70 wt % G from about 5 to about 20 wt % M from about 20 to about 40 wt %. ### Claim 2 (depends on 1) The mixture of claim 1 wherein, when analyzed by SDS-PAGE under reducing conditions and silver stained, the ratio of F.sub.1 of molecular weight approximately 48 kDa to F.sub.2 of molecular weight approximately 23 kDa is between 1:1 to about 2:1 by scanning densitometry. ### Claim 3 (depends on 1) The mixture of claim 1 which is at least about 75% pure. ### Claim 4 A mixture consisting essentially of purified fusion (F) protein, attachment (G) protein and matrix (M) protein of respiratory syncytial virus (RSV). ### Claim 5 (depends on 4) The mixture of claim 4 wherein said fusion (F) protein comprises multimeric fusion (F) proteins. ### Claim 6 (depends on 5) The mixture of claim 5 wherein, when analyzed under non-reducing conditions, said multimeric fusion (F) protein includes heterodimers of molecular weight approximately 70 kDa and dimeric and trimeric forms. ### Claim 7 (depends on 4) The mixture of claim 4 wherein, when analyzed under non-reducing conditions, said attachment (G) protein comprises G protein of molecular weight approximately 95 kDa and G protein of molecular weight approximately 55 kDa and oligomeric G protein. ### Claim 8 (depends on 4) The mixture of claim 4 wherein, when analyzed by SDS-PAGE under non-reducing conditions, said matrix (M) protein comprises M protein of molecular weight approximately 28 to 34 kDa. ### Claim 9 (depends on 4) The mixture of claim 4 wherein, when analyzed by reduced SDS-PAGE analysis, said fusion (F) protein comprises F.sub.1 of molecular weight approximately 48 kDa and F.sub.2 of molecular weight approximately 23 kDa, said attachment (G) protein comprises a G protein of molecular weight approximately 95 kDa and a G protein of molecular weight approximately 55 kDa, and said matrix (M) protein comprises an M protein of approximately 31 kDa. ### Claim 10 (depends on 4) The mixture of claim 4 which is devoid of monoclonal antibodies. ### Claim 11 (depends on 4) The mixture of claim 4 which is devoid of lentil lectin and concanavalin A. ### Claim 12 (depends on 4) The mixture of claim 4 wherein said RSV proteins are non-denatured. ### Claim 13 (depends on 4) The mixture of claim 4 wherein said RSV proteins are from one or both of subtypes RSV A and RSV B. ### Claim 14 A coisolated and copurified mixture of non-denatured proteins of respiratory syncytial virus (RSV), consisting essentially of the fusion (F) protein, attachment (G) protein and matrix (M) protein of RSV, wherein the mixture is free from lectins and is free from monoclonal antibodies. ### Claim 15 (depends on 14) The mixture of claim 14 wherein said F, G and M proteins are present in the relative proportions of: F from about 40 to about 70 wt % G from about 5 to about 20 wt % M from about 20 to about 40 wt %.