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Development and validation of novel biomarker assays for osteoarthritis

Research output: Contribution to journalArticle

Original languageEnglish
Article numbere0181334
Number of pages13
JournalPLoS ONE
Volume12
Issue number7
DOIs
DateAccepted/In press - 29 Jun 2017
DatePublished (current) - 17 Jul 2017

Abstract

BACKGROUND: Osteoarthritis (OA) is the most common chronic joint disease usually diagnosed at relatively advanced stages when there is irreparable damage to the joint(s). Recently, we have identified two novel biomarkers C3f and V65 which appear to be OA-specific and therefore potential markers of early disease. We report the development of immunoassays for quantitative measure of these two novel biomarkers.

METHOD: Monoclonal and polyclonal antibodies were generated by immunising mouse and rabbits respectively with peptide-carrier conjugates of C3f and V65. Affinity purified antibodies were used for immunoassays development and assays validated using serum from OA patients and controls.

RESULTS: The ELISAs developed showed spiked recovery of up to 96% for C3f and V65 peptides depending on serum dilutions with a coefficient of variation (CV) <10%. The intra- and inter-assay CVs for C3f and V65 were 1.3-10.8% and 4.2-10.3% respectively. Both assays were insensitive for measurements of the peptides in patients and the use of different signal amplification systems did not increase assay sensitivity.

CONCLUSION: We have developed two immunoassays for measurements of C3f and V65 peptides biomarkers discovered by our earlier proteomic study. These assays could detect the endogenous peptides in serum samples from patients and controls but lacked sensitivity for accurate measurements of the peptides in patients. Our study highlights the difficulties and challenges of validating biomarker from proteomic studies and demonstrates how to overcome some of the technical challenges associated with developing immunoassays for small peptides.

    Research areas

  • Osteoarthritis, diagnosis, biomarkers, enzyme-linked immunosorbent assay (ELISA)

    Structured keywords

  • Centre for Surgical Research

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    Rights statement: This is the final published version of the article (version of record). It first appeared online via Public Library of Science at http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0181334. Please refer to any applicable terms of use of the publisher.

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