酶联免疫吸附试验（ELISA）检测金属蛋白酶解 II 型胶原新表位（CIIM）：严重放射学骨关节炎患者血清 CIIM 升高。
Enzyme-linked immunosorbent assay (ELISAs) for metalloproteinase derived type II collagen neoepitope, CIIM--increased serum CIIM in subjects with severe radiographic osteoarthritis.
机构信息
Cartilage Biology and Biomarkers R&D, Nordic Bioscience, Herlev, Denmark.
出版信息
Clin Biochem. 2011 Apr;44(5-6):423-9. doi: 10.1016/j.clinbiochem.2011.01.001. Epub 2011 Jan 9.
OBJECTIVES
In joint degenerative diseases, the collagens are degraded by matrix metalloproteinases and protein fragments are released to serum as potential biomarkers.
METHODS
A collagen type II specific neoepitope, CIIM, was identified (…RDGAAG(1053)) by mass spectrometry. Two ELISAs against the neoepitope were developed. CIIM was measured in cartilage explants in the presence or absence of protease inhibitors. CIIM was measured in OA synovial fluid (n=51) and serum (n=156). Knee OA was graded by standard Kellgren-Lawrence (KL) score.
RESULTS
The ELISAs showed good technical performance; CV%, <13%. CIIM release from cartilage explants was blocked by the MMP inhibitor. CIIM was detected in synovial fluid. Furthermore, serum CIIM levels were significantly higher (P<0.05) in those individuals with mild or severe OA than in those with no OA.
CONCLUSION
We developed a new biomarker for joint degenerative diseases, which we demonstrated was derived from MMP-degraded type II collagen.
目的
在关节退行性疾病中，胶原蛋白被基质金属蛋白酶降解，蛋白片段作为潜在的生物标志物释放到血清中。
方法
通过质谱鉴定出一个胶原 II 型特异性新表位（CIIM）（…RDGAAG(1053)）。开发了两种针对该新表位的 ELISA 检测方法。在存在或不存在蛋白酶抑制剂的情况下，测量软骨外植体中的 CIIM。在 OA 滑液（n=51）和血清（n=156）中测量 CIIM。膝关节 OA 按标准 Kellgren-Lawrence（KL）评分进行分级。
结果
ELISA 检测方法具有良好的技术性能；CV%<13%。MMP 抑制剂可阻断软骨外植体中 CIIM 的释放。在滑液中检测到 CIIM。此外，血清 CIIM 水平在轻度或重度 OA 个体中明显高于无 OA 个体（P<0.05）。
结论
我们开发了一种用于关节退行性疾病的新型生物标志物，该标志物来源于 MMP 降解的 II 型胶原蛋白。
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