# US 8,784,827 B2 · Chimeric fusion proteins and virus like particles from Birnavirus VP2 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,784,827 B2 - Document ID: US8784827B2 - Status: Granted - Kind: B2 - Application number: 12/991,689 - Filed: 2008-05-09 - Published: 2011-08-04 - Granted: 2014-07-22 - Earliest claimed: 2008-05-09 - Assignees: Chimera Pharma S.L.U. - Applicants: Thomas Zürcher; Juan José Bernal; Cayetano von Kobbe; Ignacio Jiménez Torres; Ana Diaz Blázquez; Diana Martin Lorenzo; Gloria Calderita Lucas; Arcadio Garcia de Castro - Inventors: Thomas Zürcher; Juan José Bernal; Cayetano von Kobbe; Ignacio Jiménez Torres; Ana Diaz Blázquez; Diana Martin Lorenzo; Gloria Calderita Lucas; Arcadio Garcia de Castro - Web page: https://patentgrep.com/patent/US8784827B2 ## CPC - C07K 14/005: from viruses - A61K 39/12: Viral antigens - A61K 2039/5256: {expressing foreign proteins} - A61K 2039/5258: {Virus-like particles} - C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation - C12N 2720/10022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2720/10023: 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) - 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 field of the invention refers to chimeric Virus Like Particles (VLP) derived from Birnavirus chimeric VP2 protein. In particular, the invention refers to chimeric VP2 fusion proteins which incorporate insertions and/or substitutions with one or more amino acids or particular peptide of interest while maintaining the capacity to assemble in the form of VLP. The invention identifies particular insertion and/or substitutions sites within VP2 P loop regions and outside said P loop regions. The invention also incorporates methods for the identification of preferred insertion and substitution sites within VP2 for the incorporation of particular amino acids and peptides of interest. The resulting chimeric VLP are of interest in the design of therapeutic and prophylactic vaccines as well as in the design of drug delivery systems, carriers for DNA and RNA in gene therapy, as targeted agents, in the development of antitoxins, and as diagnostic reagents. ## Claims ### Claim 1 A fusion protein, comprising a non-naturally occuring incorporation, other than at the C- or N- terminal ends, of one or more peptides of interest, other than an Infectious Bursal Disease Virus sequence, within an Infectious Bursal Disease Virus VP2 protein, wherein the VP2 protein is selected from the group consisting of: a full length 512amino acid pVP2, a 456, 452, or 441 amino acid fragment thereof, or comprises at least 400amino acids of VP2. ### Claim 2 (depends on 1) The fusion protein according to claim 1, wherein the peptides of interest are incorporated within the VP2 P loop regions BC (Q219-G224), DE (R249-G254), FG (T283-D287) and HI (S315-Q324), and/or outside said VP2 P loop regions other than at the C- or N-terminal ends. ### Claim 3 (depends on 1) The fusion protein according to of claim 1, wherein the incorporation of peptides of interest is by insertions or substitutions of VP2 protein sequences. ### Claim 4 (depends on 3) The fusion protein according to claim 3 wherein the incorporation is by insertion. ### Claim 5 (depends on 3) The fusion protein according to claim 3 wherein the substitution by the peptide of interest replaces amino acids within VP2 P loop regions BC (Q219-G224), DE (R249-G254), FG (T283-D287) and HI (S315-Q324). ### Claim 6 (depends on 1) The fusion protein according to claim 1 wherein the peptides of interest comprise at least one lysine. ### Claim 7 (depends on 6) The fusion protein according to claim 6 wherein the one or more peptides of interest are chemically conjugated to one or more biological or chemical entities by chemical conjugation. ### Claim 8 (depends on 1) The fusion protein according to claim 1 wherein the fusion protein incorporates one or more peptides of interest at more than one site within VP2 protein at P loop regions and/or outside said VP2 P loop regions, other than at the C- or N-terminal ends. ### Claim 9 (depends on 8) The fusion protein according to claim 8 wherein the peptides of interest are the same or different for each of the insertion or substitution sites. ### Claim 10 (depends on 1) The fusion protein according to claim 1 wherein the peptide of interest is terminally fused either at its carboxy (C-) or amino (N-) terminal region, to a particular peptide of interest which may be the same or different to the peptides of interest inserted in the P loop region or outside said P loop regions. ### Claim 11 (depends on 1) The fusion protein according to claim 1, wherein fusion proteins assemble to form Virus Like Particles (VLP). ### Claim 12 (depends on 11) The fusion protein according to claim 11, wherein the Virus Like Particles present T=1 symmetry. ### Claim 13 (depends on 1) A Virus Like Particle comprising an assembly of any of the fusion proteins as defined in claim 1. ### Claim 14 (depends on 13) The Virus Like Particle comprising an assembly of the fusion proteins of claim 13 and a VP3 protein. ### Claim 15 (depends on 14) The Virus Like Particle of claim 14 in which the VP3 protein is terminally fused, either at N- or C-terminal, to a peptide of interest which may be the same or different to those incorporated in VP2. ### Claim 16 (depends on 13) The Virus Like Particle according to claim 13 which are obtained by the co-expression of pVP2 and VP3 from independent gene constructs or by the expression of constructs comprising the pVP2-VP4-VP3 polyprotein. ### Claim 17 (depends on 13) The Virus Like Particle of claim 13 forms T=13.