# US 7,407,807 B2 · Chimaeric human papillomavirus 16 I1 virus like particles and a method for preparing the particles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,407,807 B2 - Document ID: US7407807B2 - Status: Granted - Kind: B2 - Application number: 10/514,878 - Filed: 2003-05-19 - Published: 2006-02-16 - Granted: 2008-08-05 - Earliest claimed: 2002-05-17 - Assignees: University of Cape Town - Inventors: Arvind Devshi Varsani; Edward Peter Rybicki - Web page: https://patentgrep.com/patent/US7407807B2 ## CPC - C07K 14/005: from viruses - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 31/12: Antivirals - A61P 31/18: for HIV - A61P 31/20: for DNA viruses - A61P 37/04: Immunostimulants - A61K 2039/5258: {Virus-like particles} - C07K 2319/00: Fusion polypeptide - C12N 2710/20022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2710/20023: Virus like particles [VLP] - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention describes a method for producing a chimaeric human papillomavirus (HPV) L1 polypeptide containing a heterologous peptide, and in particular, a HPV L2 peptide. The method comprises the steps of introducing a DNA sequence coding for the heterologous peptide into a DNA sequence coding for the L1 polypeptide; introducing the DNA sequence including the sequences for the L1 polypeptide and heterologous peptide into a host cell in which the DNA sequence can be expressed; causing expression of the DNA sequence; and recovering the resulting chimaeric L1 polypeptide which includes the heterologous peptide. Typically, the nucleotides encoding the heterologous peptide replace the nucleotides of the L1 polypeptide at the point of insertion. The invention also describes a vector for use in the method, a host cell containing the vector, and a vaccine including the chimaeric HPV L1 polypeptide produced according to the method. ## Claims ### Claim 1 A method for producing a chimaeric polypeptide comprising human papillomavirus (HPV) L1 polypeptide containing a heterologous antigenic peptide inserted therein, the method comprising the steps of: introducing a DNA sequence encoding a heterologous antigenic peptide into a DNA sequence encoding a full-length L1 polypeptide wherein the nucleotides encoding the heterologous antigenic peptide replace a corresponding number of nucleotides within the DNA sequence encoding the L1 polypeptide thereby producing a DNA encoding a chimaeric polypeptide; and wherein said replaced nucleotides within the DNA sequence encoding the L1 polypeptide are: 241-279, 391-429, 520-558, 1240-1278 or 1291-1329 of SEQ ID NOs: 12, 6, 4, 10, and 8, respectively; introducing the DNA sequence encoding the chimaeric polypeptide into a host cell in which the DNA sequence can be expressed; causing expression of the DNA sequence; and recovering the produced chimaeric polypeptide. ### Claim 2 (depends on 1) A method according to claim 1, wherein the heterologous antigenic peptide sequence is any HPV peptide sequence which is an antigenic epitope and is B-cell specific or T-cell specific. ### Claim 3 A method for producing a chimaeric polypeptide comprising human papillomavirus (HPV) L1 polypeptide containing a HPV L2 peptide inserted therein, the method comprising the steps of: introducing a DNA sequence encoding a common-neutralizing L2 epitopic peptide into a DNA sequence encoding a full-length L1 polypeptide wherein the nucleotides encoding the common-neutralizing L2 epitopic peptide replace a corresponding number of nucleotides within the DNA sequence encoding the L1 polypeptide thereby producing a DNA encoding a chimaeric polypeptide; introducing the DNA sequence encoding the chimaeric polypeptide into a host cell in which the DNA sequence can be expressed; causing expression of the DNA sequence; and recovering the produced chimaeric polypeptide. ### Claim 4 (depends on 3) A method according to claim 3, wherein the HPV L1 polypeptide and/or HPV L2 peptide is/are HPV-16 polypeptide or peptide. ### Claim 5 (depends on 3) A method according to claim 3 wherein the HPV L2 peptide has the amino acid sequence of SEQ ID NO: 1, or a sequence which is a modification or derivative thereof, with the proviso that the modified or derived sequence has at least 80% sequence identity to the sequence of SEQ ID NO: 1 and encodes a peptide capable of eliciting an immunogenic response against HPV. ### Claim 6 (depends on 3) A method according to claim 3, wherein expression of the polypeptide is performed in either a prokaryotic or a eukaryotic expression system. ### Claim 7 (depends on 3) A method according to claim 3, wherein the chimaeric polypeptide has an amino acid sequence set out in any one of SEQ ID NOs:5, 7, 9, 11 and 13, or a sequence which has at least 80% sequence identity to any one of the amino acid sequences set out in SEQ ID NOs:5, 7, 9, 11 and 13 and which is capable of eliciting an immunogenic response against HPV. ### Claim 8 (depends on 3) A method according to claim 3, wherein the DNA sequence encoding the chimaeric polypeptide has a DNA sequence set out in any one of SEQ ID NOs: 4, 6, 8, 10 and 12, or a sequence having at least 80% sequence identity to any one of the nucleotide sequences set out in SEQ ID NOs:4, 6, 8, 10 and 12 and which encodes a chimaeric polypeptide capable of eliciting an immunogenic response against HPV. ### Claim 9 (depends on 3) A method according to claim 3, wherein the chimaeric polypeptide is capable of assembling into virus-like particles and/or capsomers. ### Claim 10 (depends on 9) A method according to claim 9, wherein the virus-like particles or capsomers are immunogenic.