Development and validation of novel biomarker assays for osteoarthritis.
<h4>Background</h4>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 pot...
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0181334&type=printable |
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| author | Khadija Ourradi Yunhe Xu Dominique de Seny John Kirwan Ashley Blom Mohammed Sharif |
| author_facet | Khadija Ourradi Yunhe Xu Dominique de Seny John Kirwan Ashley Blom Mohammed Sharif |
| author_sort | Khadija Ourradi |
| collection | DOAJ |
| description | <h4>Background</h4>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.<h4>Method</h4>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.<h4>Results</h4>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.<h4>Conclusion</h4>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. |
| format | Article |
| id | doaj-art-a08919b51ab8453096ee9d7979fdb3f1 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-a08919b51ab8453096ee9d7979fdb3f12025-08-20T03:19:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01127e018133410.1371/journal.pone.0181334Development and validation of novel biomarker assays for osteoarthritis.Khadija OurradiYunhe XuDominique de SenyJohn KirwanAshley BlomMohammed Sharif<h4>Background</h4>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.<h4>Method</h4>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.<h4>Results</h4>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.<h4>Conclusion</h4>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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0181334&type=printable |
| spellingShingle | Khadija Ourradi Yunhe Xu Dominique de Seny John Kirwan Ashley Blom Mohammed Sharif Development and validation of novel biomarker assays for osteoarthritis. PLoS ONE |
| title | Development and validation of novel biomarker assays for osteoarthritis. |
| title_full | Development and validation of novel biomarker assays for osteoarthritis. |
| title_fullStr | Development and validation of novel biomarker assays for osteoarthritis. |
| title_full_unstemmed | Development and validation of novel biomarker assays for osteoarthritis. |
| title_short | Development and validation of novel biomarker assays for osteoarthritis. |
| title_sort | development and validation of novel biomarker assays for osteoarthritis |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0181334&type=printable |
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