# US 11,078,241 B2 · Multi-epitope fusion protein of an HCV antigen and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,078,241 B2 - Document ID: US11078241B2 - Status: Granted - Kind: B2 - Application number: 16/751,609 - Filed: 2020-01-24 - Published: 2020-05-14 - Granted: 2021-08-03 - Earliest claimed: 2017-07-27 - Assignees: Roche Diagnostics Operations, Inc. - Applicants: Roche Diagnostics Operations, Inc. - Inventors: Barbara Upmeier; Ralf Bollhagen; Toralf Zarnt - Web page: https://patentgrep.com/patent/US11078241B2 ## CPC - C07K 14/005: from viruses - C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} - G01N 33/56983: {Viruses} - G01N 33/5767: {non-A, non-B hepatitis} - C07K 2319/35: containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin - C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation - C12N 2770/24222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - G01N 2333/186: Hepatitis C; Hepatitis NANB - G01N 2469/10: Detection of antigens from microorganism in sample from host - G01N 2496/00: Reference solutions for assays of biological 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) - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The disclosure relates to a multi-epitope fusion protein as well as to its use as calibrator and/or control in an in vitro diagnostics immunoassay for detecting HCV core antigen. The multi-epitope fusion protein has two to six different non-overlapping linear peptides present in the amino acid sequence of hepatitis C virus (HCV) core protein, wherein each of the peptides is separated from the other peptides by a spacer consisting of a non-HCV amino acid sequence and having a chaperone amino acid sequence. No further HCV specific amino acid sequences are present in the polypeptide. A further aspect relates to a reagent kit for detecting HCV core antigen containing said multi-epitope fusion protein as calibrator or control or both. ## Claims ### Claim 1 A multi-epitope fusion protein comprising: two to six different non-overlapping linear peptides of hepatitis C virus (HCV) core protein, wherein each of said two to six different non-overlapping linear peptides is at least 5 amino acids in length and present one to five-fold, and wherein each of said two to six different non-overlapping linear peptides is separated from the other said peptides by a spacer, wherein the spacer comprises a chaperone peptide specific of a chaperone protein, and wherein no further HCV specific amino acid sequences are present in said multi-epitope fusion protein. ### Claim 2 (depends on 1) A multi-epitope fusion protein according to claim 1, wherein said HCV core protein corresponds to an amino acid sequence of SEQ ID NO: 1. ### Claim 3 (depends on 1) A multi-epitope fusion protein according to claim 1, wherein each of said two to six linear peptides has a length of 5 to 50 amino acids. ### Claim 4 (depends on 1) A multi-epitope fusion protein according to claim 1, wherein three different linear peptides present in the amino acid sequence of said HCV core protein are part of said multi-epitope fusion protein. ### Claim 5 (depends on 1) A multi-epitope fusion protein according to claim 1, wherein said three different linear peptides comprises a linear amino acid sequence present within amino acid positions 55 to 80 of SEQ ID NO:1, amino acid positions 90 to 120 of SEQ ID NO:1 and amino acid positions 145 to 175 of SEQ ID NO:1. ### Claim 6 (depends on 1) A multi-epitope fusion protein according to claim 1, having the formula X-spacer-Y-spacer-Z wherein X, Y and Z designate different non-overlapping linear peptides of HCV core protein; wherein Z is present one to five-fold and wherein spacer designates a chaperone peptide. ### Claim 7 (depends on 6) A multi-epitope fusion protein according to claim 6, wherein X comprises a linear amino acid sequence present within amino acid positions 55 to 80 of SEQ ID NO:1, Y comprises linear amino acid sequence present within amino acid positions 90 to 120 of SEQ ID NO:1, Z comprises linear amino acid sequences present within amino acid positions 145 to 175 of SEQ ID NO:1. ### Claim 8 (depends on 1) A multi-epitope fusion protein according to claim 1, wherein said spacer comprises a sequence of at least 25 non-HCV amino acids. ### Claim 9 (depends on 1) A multi-epitope fusion protein according to claim 1, consisting of any of SEQ ID NOs:4 to 6. ### Claim 10 (depends on 1) A method of producing a soluble multi-epitope fusion protein, said method comprising the steps of a) culturing host cells transformed with an expression vector comprising operably linked a recombinant DNA molecule encoding a multi-epitope fusion protein according to claim 1, b) expression of said polypeptide and c) purification of said polypeptide. ### Claim 11 (depends on 1) A reagent kit for the detection of HCV core antigen, comprising in separate containers or in separated compartments of a single container at least one HCV core antigen-specific binding agent and at least one multi-epitope fusion protein according to claim 1 as calibrator material. ### Claim 12 (depends on 11) A reagent kit according to claim 11, comprising at least two HCV core antigen-specific binding agents, wherein one of said at least two HCV core antigen-specific binding agents is detectably labeled. ### Claim 13 (depends on 11) A reagent kit according to claim 11, additionally comprising microparticles.