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However, serum or plasma CTX-II have not been demonstrated to have any clinical utility to date. Here, we describe the GPDPLQ<jats:sub>1237<\/jats:sub> ELISA which targets the EKGPDPLQ\u2193 neo-epitope, an elongated version of the CTX-II neo-epitope (EKGPDP\u2193), speculated to be a blood-precursor of CTX-II generated by the cysteine protease cathepsin K. Human osteoclast cartilage resorption cultures as well as oncostatin M and tumour necrosis factor \u03b1-stimulated bovine cartilage explant cultures were used to validate GPDPLQ<jats:sub>1237<\/jats:sub> biologically by treating the cultures with the cysteine protease inhibitor E-64 and\/or the matrix metalloproteinase (MMP) inhibitor GM6001 to assess the potential contributions of these two protease classes to GPDPLQ<jats:sub>1237<\/jats:sub> release. Cartilage resorption-derived GPDPLQ<jats:sub>1237<\/jats:sub> release was inhibited by E-64 (72.1% inhibition), GM6001 (75.5%), and E-64\/GM6001 (91.5%), whereas CTX-II release was inhibited by GM6001 (87.0%) but not by E-64 (5.5%). Cartilage explant GPDPLQ<jats:sub>1237<\/jats:sub> and CTX-II release were both fully inhibited by GM6001 but were not inhibited by E-64. No clinically relevant GPDPLQ<jats:sub>1237<\/jats:sub> reactivity was identified in human serum, plasma, or urine from healthy donors or arthritis patients. In conclusion, the GPDPLQ<jats:sub>1237<\/jats:sub> biomarker is released during osteoclast-derived cysteine protease- and MMP-mediated cartilage degradation <jats:italic>in vitro<\/jats:italic>, whereas CTX-II release is mediated by MMPs and not by cysteine proteases, as well as from MMP-mediated cartilage degradation under a pro-inflammatory stimulus. These findings suggest that GPDPLQ<jats:sub>1237<\/jats:sub> may be relevant in diseases with pathological osteoclast activity and cartilage degradation. Further studies are required to validate the neo-epitope in human samples.<\/jats:p>","DOI":"10.1038\/s41598-019-39803-0","type":"journal-article","created":{"date-parts":[[2019,2,28]],"date-time":"2019-02-28T11:05:03Z","timestamp":1551351903000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["GPDPLQ1237\u2014A Type II Collagen Neo-Epitope Biomarker of Osteoclast- and Inflammation-Derived Cartilage Degradation in vitro"],"prefix":"10.1038","volume":"9","author":[{"given":"Henrik","family":"L\u00f6fvall","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Katri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aneta","family":"D\u0105browska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Morten A.","family":"Karsdal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunyun","family":"Luo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi","family":"He","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tina","family":"Manon-Jensen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Morten H.","family":"Dziegiel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne-Christine","family":"Bay-Jensen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian S.","family":"Thudium","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kim","family":"Henriksen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,2,28]]},"reference":[{"key":"39803_CR1","doi-asserted-by":"publisher","first-page":"683","DOI":"10.1016\/j.joca.2014.02.007","volume":"22","author":"AM Valdes","year":"2014","unstructured":"Valdes, A. 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