Diaz Romero J, Outschoorn I
Servicio de Inmunologia, Centro Nacional de Microbiologia (C.N.M.V.I.S), Majadahonda, Madrid, Spain.
J Immunol Methods. 1993 Mar 15;160(1):35-47. doi: 10.1016/0022-1759(93)90006-s.
A method is described for the selective biotinylation of meningococcal capsular polysaccharide from Neisseria meningitidis group B and its application to an enzyme-linked immunoabsorbent assay (ELISA) to detect specific antibodies by immobilization on streptavidin-coated microtiter wells. Capsular polysaccharide from Neisseria meningitidis B has been biotinylated by specific periodate oxidation of terminal residues and condensation of the resulting aldehydes with biotin hydrazide, using a spin-column technique in the intermediate purification steps. The ELISA was optimized employing an extended reaction time between the label alkaline phosphatase and its most common substrate, p-nitrophenyl phosphate, together with evaluation of blocking agents to minimize non-specific binding. Specificity was demonstrated by a direct competitive enzyme immunoassay (EIA).
本文描述了一种从B群脑膜炎奈瑟菌中选择性生物素化脑膜炎球菌荚膜多糖的方法及其在酶联免疫吸附测定（ELISA）中的应用，该方法通过将其固定在包被抗生物素蛋白的微量滴定孔上来检测特异性抗体。B群脑膜炎奈瑟菌的荚膜多糖已通过末端残基的特定高碘酸盐氧化以及所得醛与生物素酰肼的缩合进行生物素化，在中间纯化步骤中使用了旋转柱技术。ELISA通过延长标记碱性磷酸酶与其最常见底物对硝基苯磷酸之间的反应时间以及评估封闭剂以最小化非特异性结合进行了优化。通过直接竞争酶免疫测定（EIA）证明了其特异性。
