杀虫剂西维因单克隆抗体的研制及鉴定

Preparation and identification of monoclonal antibodies for insecticide carbaryl

目的 制备特异性的西维因单克隆抗体。方法 合成了西维因的半抗原并对其进行了结构鉴定 ,将半抗原与牛血清白蛋白和卵清蛋白共价偶联分别制备免疫抗原和包被抗原。再将免疫的Balb c小鼠脾脏细胞与SP2 0小鼠骨髓瘤细胞融合 ,建立一株分泌西维因单克隆抗体的杂交瘤细胞。分析该杂交瘤细胞的染色体 ,并以间接酶联免疫吸附分析法 (ELISA)检测了单克隆抗体免疫球蛋白的类型及其与西维因的亲和性和特异性。结果 杂交瘤细胞的平均染色体数目为 98条 ,它分泌IgG1亚类的单克隆抗体 ;该单克隆抗体与西维因的亲和性较高 (IC50 =5 2ng mL)而与西维因结构类似物的交叉反应很小。结论 本实验成功的制备了西维因特异性的单克隆抗体 ,为其ELISA方法的建立提供了条件。

Objective To prepare monoclonal antibodies for insecticide carbaryl. Methods Hapten of carbaryl was synthesized and the molecule structure was identified. The hapten was covalently conjugated to bovine sera albumin (BSA) and ovalbumin (OVA) to prepare immunogen and coating antigen, respectively. One hybridoma cell line secreting monoclonal antibodies (McAb) against carbaryl has been established by fusing mouse myeloma cells SP2/0 and splenocytes from Balb/c mice immunized with immunogen. The chromosomes of hybridoma cell were analyzed and the immunoglobulin class, affinity and specificity of McAb were determinated with indirect enzyme linked immunosorbent assay (ELISA). Results The average of chromosomes in a hybridoma cell was 98 and the immunoglobulin class of McAb was IgG1. McAbs showed a high affinity for carbaryl (IC 50 =52 ng/mL) and minimum cross reactivity for several analogous compounds of carbaryl. Conclusion The specific monoclonal antibodies of carbaryl were obtained successfully in this experiment and this made it possible to establish ELISA for carbaryl.