Kongtawelert P, Ghosh P
Raymond Purves Research Laboratories, (University of Sydney, Royal North Shore Hospital of Sydney, St. Leonards, N.S.W., Australia.
Clin Chim Acta. 1990 Dec 31;195(1-2):17-26. doi: 10.1016/0009-8981(90)90190-4.
A new sandwich-ELISA for the determination of keratan sulphate peptides in biological fluids is described. The technique involves binding a commercially available monoclonal antibody against keratan sulphate to microtitre plates, adding the unknown keratan sulphate antigen then interacting the keratan sulphate-antibody complex with a biotin-monoclonal antibody conjugate which was also specific for keratan sulphate peptides. Alkaline phosphatase conjugated streptravidin was then added and the amount which bound to the biotin was determined by measuring the release of chromogen from an added chromogenic substrate. Using this assay, keratan sulphate peptides in biological fluids within the range 10-1000 ng/ml could be quantitated. This method was found to be more sensitive than presently used techniques. The intra- and inter-assay coefficients of variation were 11% and 13%, respectively.
本文描述了一种用于测定生物体液中硫酸角质素肽的新型夹心酶联免疫吸附测定法（sandwich-ELISA）。该技术包括将一种市售的抗硫酸角质素单克隆抗体结合到微量滴定板上，加入未知的硫酸角质素抗原，然后使硫酸角质素 - 抗体复合物与同样对硫酸角质素肽具有特异性的生物素 - 单克隆抗体缀合物相互作用。接着加入碱性磷酸酶偶联的链霉亲和素，并通过测量从添加的显色底物中释放的显色剂来确定与生物素结合的量。使用该测定法，可以对浓度范围在10 - 1000 ng/ml的生物体液中的硫酸角质素肽进行定量。结果发现该方法比目前使用的技术更灵敏。测定内和测定间的变异系数分别为11%和13%。
