Heimer R
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Anal Biochem. 1989 Aug 1;180(2):211-5. doi: 10.1016/0003-2697(89)90117-6.
Using synovial fluid of individuals with osteoarthritis as a prototypic biologic fluid containing one part proteoglycan per 100 to 1000 parts of other protein, profiles of proteoglycan were produced without preliminary purification through agarose-acrylamide gel electrophoresis, transfer to nitrocellulose, and triple immunoblotting. Chondroitin sulfate, keratan sulfate, and a hyaluronic acid binding region appear to be present on individual synovial fluid proteoglycans in variable amounts, and consequently a triple immunoblot using monoclonal antibodies to these three epitopes has the potential for developing a proteoglycan profile. The profile is assembled by means of densitometric scans of autoradiograms obtained after use of 125I-labeled anti-mouse immunoglobulin. By contrast to the profile of a relatively homogeneous proteoglycan purified from articular cartilage extracts, the proteoglycans of synovial fluid appeared to be quite heterogeneous with the bulk of keratan sulfate epitopes migrating ahead of the bulk of the chondroitin sulfate epitopes. Most of the proteoglycans appeared to possess a hyaluronate binding region.
以骨关节炎患者的滑液作为一种典型的生物流体，其中蛋白聚糖与其他蛋白质的比例为每100至1000份含1份，通过琼脂糖-丙烯酰胺凝胶电泳、转印至硝酸纤维素膜以及三重免疫印迹法，无需预先纯化即可生成蛋白聚糖谱。硫酸软骨素、硫酸角质素和透明质酸结合区域似乎以不同含量存在于单个滑液蛋白聚糖上，因此使用针对这三个表位的单克隆抗体进行三重免疫印迹法有潜力生成蛋白聚糖谱。该谱是通过对使用125I标记的抗小鼠免疫球蛋白后获得的放射自显影片进行光密度扫描来组装的。与从关节软骨提取物中纯化得到的相对均质的蛋白聚糖谱相比，滑液中的蛋白聚糖显得相当不均一，大部分硫酸角质素表位迁移至大部分硫酸软骨素表位之前。大多数蛋白聚糖似乎都具有透明质酸结合区域。
