# US 7,425,437 B2 · Vaccines against West Nile Virus > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,425,437 B2 - Document ID: US7425437B2 - Status: Granted - Kind: B2 - Application number: 11/110,517 - Filed: 2005-04-20 - Published: 2005-08-25 - Granted: 2008-09-16 - Earliest claimed: 1999-11-26 - Assignees: Crucell Holland B.V. - Inventors: Alphonsus G. C. M. UytdeHaag; Govert Johan Schouten; Jaap Goudsmit - Web page: https://patentgrep.com/patent/US7425437B2 ## CPC - A61K 39/12: Viral antigens - 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) - A61K 2039/5252: {inactivated (killed)} - A61K 2039/552: {Veterinary vaccine} - A61K 2039/55505: {Inorganic adjuvants} - A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59} - C12N 2770/24134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2770/24163: by chemical treatment - 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) ## Abstract The present invention relates to novel vaccines containing (whole-inactivated) West Nile Viruses and/or West Nile viral proteins derived therefrom, produced on human cells, wherein the human cells comprise a sequence encoding at least one early region-1 (E1) gene product of an adenovirus. The cells are preferably cultured in suspension to very high densities and under serum-free conditions. Herein, it is disclosed that use of such cells results in high titers of West Nile Virus produced. ## Claims ### Claim 1 A method of producing a West Nile Virus, said method comprising: infecting a cell with West Nile Virus, wherein the cell comprises an E1 region of an adenovirus integrated into the cell's chromosomal genome; culturing the cell in a suitable medium under conditions that allow for the production of the West Nile Virus; and producing West Nile Virus. ### Claim 2 (depends on 1) The method according to claim 1, wherein the cell is a non-tumorous human cell. ### Claim 3 (depends on 1) The method according to claim 1, wherein the cell is a retinoblast. ### Claim 4 (depends on 1) The method according to claim 1, wherein the cell is an embryonic cell. ### Claim 5 (depends on 1) The method according to claim 1, wherein the cell is an embryonic retinoblast. ### Claim 6 (depends on 1) The method according to claim 1, wherein the cell is a PER.C6® cell. ### Claim 7 (depends on 1) The method according to claim 1, wherein the cell is a kidney cell or an ammocyte. ### Claim 8 (depends on 1) The method according to claim 1, wherein said West Nile Virus is selected from the group consisting of lineage II strain West Nile B956, lineage II strain Madagascar 1978, lineage II strain Cyprus 1968, lineage I strain Kunjin 1960, lineage I strain Kunjin 1991, lineage I strain Goose Israel 1998, and lineage I strain New York 1999. ### Claim 9 (depends on 1) The method according to claim 1, wherein said West Nile Virus is a lineage II strain selected from the group consisting of Kenya, Uganda, Senegal 1990, Uganda 1937, Uganda 1959, Central African Republic 1972a, Central African Republic 1972b, Central African Republic 1983, Madagascar 1986 and Madagascar 1988. ### Claim 10 (depends on 1) The method according to claim 1, wherein said cell is infected with a West Nile Virus at a multiplicity of infection ranging from about 5 to about 5×10−7 plaque forming units per cell. ### Claim 11 (depends on 1) The method according to claim 1, wherein culturing the cell comprises culturing the cell in serum-free medium. ### Claim 12 (depends on 1) The method according to claim 1, wherein culturing the cell comprises culturing the cell in a suspension culture. ### Claim 13 (depends on 1) The method according to claim 1, wherein the cell comprises a temperature-sensitive E2A gene. ### Claim 14 (depends on 1) The method according to claim 1, further comprising inactivating the produced West Nile Virus. ### Claim 15 (depends on 14) The method according to claim 14, further comprising: harvesting the produced West Nile Virus, wherein harvesting comprises purifying the West Nile Virus particles. ### Claim 16 (depends on 1) The method according to claim 1, further comprising: disrupting the produced West Nile Virus; and purifying one or more anti genie components of said West Nile Virus. ### Claim 17 (depends on 14) The method according to claim 14, wherein said produced West Nile Virus is inactivated with formaldehyde.