Møller H J, Møller-Pedersen T, Damsgaard T E, Poulsen J H
Department of Clinical Biochemistry, K.H. University Hospital, Aarhus, Denmark.
Clin Chim Acta. 1995 May 15;236(2):195-204. doi: 10.1016/0009-8981(95)06053-g.
The prototype monoclonal keratan sulphate (KS) antibody 5D4 that is widely used for detection of KS in tissues and biological fluids reacts strongly with commercial low grade shark cartilage chondroitin 6-sulphate. Characterization of the immunogenic material by chondroitinase ABC digestion, ELISA inhibition studies, immunoblotting and HPLC analyses confirmed the presence of substantial amounts of KS, probably as a large proteoglycan (> 120 kDa). Commercial and heterogenic glycosaminoglycan preparations therefore must be used with great caution in immunological analyses. On the other hand the shark cartilage chondroitin 6-sulphate is an easy accessible source of immunogenic KS that can be used as a reference standard and as coating antigen in KS-ELISAs. The concentration of immunogenic KS in synovial fluid measured with an ELISA based solely on reagents of shark cartilage chondroitin 6-sulphate correlated well (r = 0.90) with the concentrations obtained with a traditional KS-ELISA that uses purified aggrecan as standard and coating antigen, and KS in both serum and synovial fluid could be measured with sufficient linearity.
广泛用于检测组织和生物体液中硫酸角质素（KS）的原型单克隆抗体5D4与市售低级别鲨鱼软骨硫酸软骨素6-硫酸盐发生强烈反应。通过软骨素酶ABC消化、ELISA抑制研究、免疫印迹和HPLC分析对免疫原性物质进行表征，证实存在大量的KS，可能是以一种大分子蛋白聚糖（> 120 kDa）的形式存在。因此，在免疫分析中必须极其谨慎地使用市售和异质性糖胺聚糖制剂。另一方面，鲨鱼软骨硫酸软骨素6-硫酸盐是一种易于获取的免疫原性KS来源，可作为参考标准以及KS-ELISA中的包被抗原。仅基于鲨鱼软骨硫酸软骨素6-硫酸盐试剂的ELISA所测得的滑液中免疫原性KS浓度与使用纯化聚集蛋白聚糖作为标准和包被抗原的传统KS-ELISA所测得的浓度具有良好的相关性（r = 0.90），并且血清和滑液中的KS均可实现足够的线性测量。
