# US 8,772,043 B2 · Universal peptide tags for transgene polypeptide analysis by mass spectrometry > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,772,043 B2 - Document ID: US8772043B2 - Status: Granted - Kind: B2 - Application number: 13/334,420 - Filed: 2011-12-22 - Published: 2012-06-28 - Granted: 2014-07-08 - Earliest claimed: 2010-12-23 - Assignees: Pioneer Hi-Bred International, Inc. - Applicants: Xuehua Hu; Michaela Owens; Clara Alarcon; Xiping Niu - Inventors: Xuehua Hu; Michaela Owens; Clara Alarcon; Xiping Niu - Web page: https://patentgrep.com/patent/US8772043B2 ## CPC - C12N 9/1029: {transferring groups other than amino-acyl groups (2.3.1)} - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - G01N 33/6848: {Methods of protein analysis involving mass spectrometry} - G01N 33/6851: {Methods of protein analysis involving laser desorption ionisation mass spectrometry} - A01K 2227/105: Murine - A01K 2267/01: Animal expressing industrially exogenous proteins - C07K 2319/00: Fusion polypeptide - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS ## Abstract Compositions and methods that allow for the rapid detection and accurate quantification of any polypeptides of interest are provided. Compositions include isolated polypeptides comprising at least one universal peptide tag, as well as isolated polynucleotides encoding such polypeptides. The universal peptide tags can be quantified by methods including, but not limited to, mass spectrometry, and can act as surrogates for determining the concentration of the polypeptides comprising the universal peptide tags. Methods provide for the detection and/or quantification of any polypeptides of interest that comprise at least one universal peptide tag, including methods using mass spectroscopy techniques. Methods are also provided for producing hosts, or cells or parts thereof, that comprise polypeptides comprising at least one universal peptide tag. Hosts, or cells, or parts thereof, include mammalian, bacterial, insect, yeast, viral or plant. ## Claims ### Claim 1 A method of determining the concentration of a polypeptide in a cell or part thereof, wherein said polypeptide comprises at least one universal peptide tag that is incorporated into the amino acid sequence of said polypeptide, and wherein said polypeptide does not comprise an epitope tag, said method comprising: a. analyzing a sample for the presence of said at least one universal peptide tag in said host, or cell or part thereof, wherein said analysis is performed by mass spectrometry, and further wherein said at least one universal peptide tag remains intact; b. detecting the presence of said polypeptide based on the presence of said at least one universal peptide tag in said sample; c. determining the concentration of said at least one universal peptide tag in said cell, or part thereof; and, d. calculating the concentration of said polypeptide based on the concentration of said at least one universal peptide tag in said sample. ### Claim 2 (depends on 1) The method of claim 1, wherein said at least one universal peptide tag comprises the amino acid sequence set forth in SEQ ID NO: 1, or an amino acid sequence having one substitution when compared to the amino acid sequence set forth in SEQ ID NO: 1, wherein said at least one universal peptide tag can be detected by mass spectrometry and can be used to determine the presence or concentration of said polypeptide in a sample. ### Claim 3 (depends on 1) The method of claim 1, wherein said at least one universal peptide tag is fused to the N-terminus of the polypeptide, is fused to the C-terminus of the polypeptide, or is incorporated into the amino acid sequence of the polypeptide and is not located at the N-terminus or C-terminus. ### Claim 4 (depends on 1) The method of claim 1, wherein a linker peptide is positioned between said at least one universal peptide tag and the amino acid sequence of said polypeptide. ### Claim 5 (depends on 4) The method of claim 4, wherein said linker peptide comprises an amino acid sequence set forth in SEQ ID NO: 13. ### Claim 6 (depends on 1) The method of claim 1, wherein said polypeptide is encoded by a polynucleotide comprising a nucleotide sequence, wherein said nucleotide sequence encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1, or an amino acid sequence having one substitution when compared to the amino acid sequence set forth in SEQ ID NO: 1. ### Claim 7 (depends on 1) The method of claim 1, wherein said cell is a bacteria, a virus, or a yeast. ### Claim 8 (depends on 2) The method of claim 2, wherein said at least one universal peptide tag comprises the amino acid sequence set forth in SEQ ID NO:1. ### Claim 9 (depends on 6) The method of claim 6, wherein said nucleotide sequence encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 1. ### Claim 10 (depends on 6) The method of claim 6, wherein said nucleotide sequence comprises SEQ ID NO: 2.