Verschoor J A, Meiring M J, van Wyngaardt S, Weyer K
Department of Biochemistry, University of Pretoria, Republic of South Africa.
J Immunoassay. 1990;11(4):413-28. doi: 10.1080/01971529008055042.
Several methods of coating whole cells of Mycobacterium tuberculosis H37 RV to ELISA microtitre plates were compared with the aim of developing an ELISA screening assay for murine monoclonal antibodies in culture supernatants and human antibodies in patient sera. Undercoats of nylon or poly-L-lysine were compared to polystyrene as adsorptive surfaces for the bacteria, the effect of increased ionic strength and iclusion of SDS in the coating buffer measured, and methanol (70%) and glutaraldehyde (5%) investigated for their efficiency as fixatives of the bacterial monolayers. The results suggest PBS as a satisfactory coating buffer for the bacterial cells on polystyrene, and 70% methanol the preferred fixative for the dried antigen-coated plates.
比较了几种将结核分枝杆菌H37 RV全细胞包被到ELISA微量滴定板上的方法,目的是开发一种用于检测培养上清液中鼠单克隆抗体和患者血清中人抗体的ELISA筛选试验。将尼龙或聚-L-赖氨酸底涂层与聚苯乙烯作为细菌的吸附表面进行比较,测量了涂层缓冲液中离子强度增加和SDS加入的效果,并研究了甲醇(70%)和戊二醛(5%)作为细菌单层固定剂的效率。结果表明,PBS是聚苯乙烯上细菌细胞的令人满意的包被缓冲液,70%甲醇是干燥抗原包被板的首选固定剂。