Verschoor J A, Vermeulen N M, Visser L
Department of Biochemistry, University of Pretoria, Republic of South Africa.
J Immunol Methods. 1990 Feb 20;127(1):43-9. doi: 10.1016/0022-1759(90)90339-w.
An ELISA system, based on the novel use of a hapten-nylon conjugate as solid-phase coating antigen, has been applied in the screening of hybridoma cultures for anti-hapten monoclonal antibodies directed against the herbicide atrazine and its derivatives. Conjugation of a 2-aminocaproic acid derivative of atrazine with DCC to polyamide (Nylon 6) gave haptenated nylon which was soluble in aqueous cresol-ethanol mixtures and adsorbed efficiently on polystyrene microtitre plates. Reproducible ELISA results were obtained with culture supernatants of hybridomas derived from spleen cells of mice that had been immunized with atrazine-bovine serum albumin conjugates. Satisfactory results were also obtained with a water soluble peptide conjugated to nylon for use as a coating antigen in an ELISA. Plates coated with hapten-nylon as antigen have the added advantage that they can be stored at room temperature for at least 6 months without loss of activity. Nylon therefore appears to have general applicability as a carrier for both non-polar and polar haptens in the preparation and use of coating antigens.
一种基于将半抗原 - 尼龙缀合物用作固相包被抗原的新型应用的酶联免疫吸附测定（ELISA）系统，已被应用于筛选杂交瘤培养物，以获取针对除草剂阿特拉津及其衍生物的抗半抗原单克隆抗体。阿特拉津的2 - 氨基己酸衍生物与二环己基碳二亚胺（DCC）与聚酰胺（尼龙6）共轭，得到半抗原化的尼龙，其可溶于甲酚 - 乙醇混合水溶液，并能有效吸附在聚苯乙烯微量滴定板上。用来自用阿特拉津 - 牛血清白蛋白缀合物免疫的小鼠脾细胞产生的杂交瘤培养上清液获得了可重复的ELISA结果。用与尼龙共轭的水溶性肽作为ELISA中的包被抗原也获得了满意的结果。用半抗原 - 尼龙作为抗原包被的平板具有额外的优点，即它们可以在室温下储存至少6个月而不会丧失活性。因此，尼龙似乎在包被抗原的制备和使用中作为非极性和极性半抗原的载体具有普遍适用性。
