# US 2010/0168402 A1 · Direct Attachment of Polypeptides to Virus Like Particles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2010/0168402 A1 - Document ID: US20100168402A1 - Status: Published application - Kind: A1 - Application number: 12/619,442 - Filed: 2009-11-16 - Published: 2010-07-01 - Earliest claimed: 2008-11-14 - Inventors: Bradley C. Bundy; James R. Swartz - Web page: https://patentgrep.com/patent/US20100168402A1 ## CPC - C07K 14/005: from viruses - 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 2039/5258: {Virus-like particles} - C12N 2795/18122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2795/18123: 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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 Compositions and methods are provided for the control of direct protein attachment to virus like particles where virus structural proteins that have been modified to comprise an unnatural amino acid at a pre-determined site are reacted with one or more “display” polypeptides that also comprise an unnatural amino acid at a pre-determined site in a one step reaction. The compositions of the invention are useful for various purposes where it is desirable to efficiently and directly attach multiple polypeptides to a single carrier entity, particularly where two or more different polypeptides are attached to a single carrier. ## Claims ### Claim 1 A method for direct assembly of display polypeptide-modified virus like particles, the method comprising: combining in a reaction mix: (a) a stable virus like particle (VLP) free of a viral genome comprising carrier virus coat polypeptides modified to comprise at least one first unnatural amino acid at a pre-determined site with (b) display polypeptides modified to comprise at least one second unnatural amino acid, wherein the first unnatural amino acid is different from, and reactive with, the second unnatural amino acid; reacting the first and second unnatural amino acids in a single step to form a stable attachment; wherein the VLP comprises at least 60 display polypeptides. ### Claim 2 (depends on 1) The method of claim 1, wherein said carrier virus coat polypeptides comprise less than three unnatural amino acids in each polypeptide. ### Claim 3 (depends on 1) The method of claim 1, wherein said carrier virus coat polypeptides comprise a single unnatural amino acid in each polypeptide. ### Claim 4 (depends on 1) The method of claim 1, wherein said display polypeptides comprise less than three unnatural amino acids in each polypeptide. ### Claim 5 (depends on 1) The method of claim 1, wherein said display polypeptides comprise a single unnatural amino acid in each polypeptide. ### Claim 6 (depends on 1) The method of claim 1, wherein said first and said second unnatural amino acids are selected from p-acetyl-L-phenylalanine, p-propargyloxyphenylalanine, and p-azido-L-phenylalanine. ### Claim 7 (depends on 6) The method of claim 6, wherein said first and said second unnatural amino acids are p-propargyloxyphenylalanine and p-azido-L-phenylalanine. ### Claim 8 (depends on 7) The method of claim 7, wherein said reacting is performed in the presence of a Cu(I) catalyst. ### Claim 9 (depends on 1) The method of claim 1, wherein following said reacting step, at least 50% of the unnatural amino acids present on the carrier virus coat polypeptides are stably attached to display polypeptide. ### Claim 10 (depends on 9) The method of claim 9, wherein following said reacting step, at least 75% of the unnatural amino acids present on the carrier virus coat polypeptides are stably attached to display polypeptide. ### Claim 11 (depends on 1) The method of claim 1, wherein two or more display polypeptides are present in the reaction mix, therein providing for a display polypeptide-modified virus like particle comprising said two or more display polypeptides. ### Claim 12 (depends on 1) The method of claim 1, wherein said display polypeptides are greater than 15 amino acids in length. ### Claim 13 (depends on 1) The method of claim 1, wherein said display polypeptides comprise said second unnatural amino acid at a position other than the amino terminus. ### Claim 14 (depends on 1) A kit for use the method according to claim 1. ### Claim 15 (depends on 1) A display polypeptide-modified virus like particle produced by a method set forth in claim 1.