Department of Orthopedic Surgery, Cell Therapy Center, Ajou University, Suwon 443-749, Korea.
J Microbiol Biotechnol. 2012 May;22(5):654-8. doi: 10.4014/jmb.1111.11005.
Osteoarthritis (OA) is the most common rheumatic pathology. One of the major objectives of OA research is the development of early diagnostic strategies such as those using proteomic technology. Synovial fluid (SF) in OA patients is a potential source of biomarkers for OA. The efficient and reliable preparation of SF proteomes is a critical step towards biomarker discovery. In this study, we have optimized a pretreatment method for two-dimensional gel electrophoresis (2DE) separation of the SF proteome, by enriching low-abundance proteins and simultaneously removing hyaluronic acid, albumin, and IgG. SF samples pretreated using this optimized method were then evaluated by 1DE and 2DE separation followed by immunodetection of cartilage oligomeric matrix protein (COMP), a known OA biomarker, and by the identification of 3 proteins (apolipoprotein, haptoglobin precursor, and fibrinogen D fragment) that are related to joint diseases.
骨关节炎 (OA) 是最常见的风湿性病理。OA 研究的主要目标之一是开发早期诊断策略，如使用蛋白质组学技术。OA 患者的滑液 (SF) 是 OA 生物标志物的潜在来源。SF 蛋白质组的高效可靠制备是发现生物标志物的关键步骤。在这项研究中，我们通过富集低丰度蛋白质并同时去除透明质酸、白蛋白和 IgG，优化了用于 SF 蛋白质组二维凝胶电泳 (2DE) 分离的预处理方法。然后，通过 1DE 和 2DE 分离以及对已知 OA 生物标志物软骨寡聚基质蛋白 (COMP) 的免疫检测，以及对与关节疾病相关的 3 种蛋白质 (载脂蛋白、触珠蛋白前体和纤维蛋白原 D 片段) 的鉴定，评估了使用这种优化方法预处理的 SF 样本。
