Citrullinated peptides for diagnosing and prognosing rheumatoid arthritis
View Patent ↗The present invention provides novel citrullinated peptides, their use in methods for aiding, assisting, improving, or facilitating the diagnosis or prognosis of rheumatic diseases such as rheumatoid arthritis (RA), and methods for identifying novel citrullinated peptides that are immunoreactive with anti-citrullinated protein antibodies (ACPAs). The present invention also provides methods for detecting rheumatoid factor (RF) using novel RF detection reagents as a means to aid, assist, improve, or facilitate the diagnosis or prognosis of rheumatic diseases such as RA. Kits comprising at least one of the novel citrullinated peptides and/or RF detection reagents of the present invention are also provided.
1. A synthetic peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, and 67, wherein said peptide is immunologically reactive with an anti-citrullinated protein antibody.
2. The synthetic peptide of claim 1 , wherein said peptide further comprises a tag.
3. The synthetic peptide of claim 2 , wherein said tag is biotin.
4. A synthetic peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, and 66 wherein said peptide is immunologically reactive with an anti-citrullinated protein antibody.
5. The synthetic peptide of claim 1 or 4 , wherein the C-terminus of said peptide is amidated.
6. The synthetic peptide of claim 1 or 4 , wherein said peptide further comprises a detectable moiety.
7. The synthetic peptide of claim 6 , wherein said detectable moiety is a fluorescent moiety.
8. A method for detecting an anti-citrullinated protein antibody in a biological sample, the method comprising the steps of:
(a) contacting the biological sample with a synthetic peptide of claim 1 or 4 under conditions suitable to transform said peptide into a complex comprising said peptide and anti-citrullinated protein antibody; and
(b) detecting the presence or level of said complex.
9. The method of claim 8 , wherein step (b) comprises the detection of a detectable moiety conjugated to said peptide.
10. The method of claim 9 , wherein said detectable moiety is a fluorescent moiety.
11. The method of claim 8 , wherein step (b) comprises the sub-steps of:
(i) contacting said complex with a detection reagent comprising a reporter group to transform said complex into a labeled complex; and
(ii) detecting the presence or level of said labeled complex.
12. The method of claim 11 , wherein said detection reagent is selected from the group consisting of an anti-IgA antibody, an anti-IgG antibody, an anti-IgM antibody, Protein L, Protein A, Protein G, and mixtures thereof.
13. The method of claim 11 , wherein said reporter group is selected from the group consisting of radioactive groups, fluorescent groups, luminescent groups, enzymes, biotin, and dyes.
14. The method of claim 11 , wherein detecting the presence or level of said labeled complex comprises detecting the presence or level of a signal generated from said reporter group.
15. The method of claim 11 , wherein said anti-citrullinated protein antibodies comprise IgG anti-citrullinated protein antibodies.
16. A kit comprising:
(a) at least one synthetic peptide of claim 1 or 4 ; and
(b) a detection reagent comprising a detectable moiety.
17. A kit comprising:
(a) at least one synthetic peptide of claim 1 or 4 ; and
(b) a detectable moiety linked to said at least one synthetic peptide.
18. The kit of claim 16 or 17 , wherein said detectable moiety is selected from the group consisting of radioactive groups, fluorescent groups, luminescent groups, enzymes, biotin, and dyes.
19. The kit of claim 16 , wherein said detection reagent is selected from the group consisting of an anti-IgA antibody, an anti-IgG antibody, an anti-IgM antibody, Protein L, Protein A, Protein G, and mixtures thereof.
20. The kit of claim 16 or 17 , wherein said at least one synthetic peptide is immobilized on a solid support.