溴氰菊酯单克隆抗体的制备及初步应用

Preparation and preliminary application of monoclonal antibody against deltamethrin

目的制备溴氰菊酯单克隆抗体。方法用席夫碱反应合成的溴氰菊酯偶联牛血清白蛋白全抗原免疫BALB／c小鼠，采用杂交瘤技术获得溴氰菊酯的单克隆抗体。利用酶联免疫吸附分析、十二烷基磺酸钠聚丙烯酰胺凝胶电泳等技术对纯化后的抗体进行表征。结果制得抗体为lgG1亚型；SY子量为168kDa，重链为60kDa，轻链为24kDa；亲和常数为4．75×108L／mol；该抗体为特异性的。利用间接竞争酶联免疫吸附法对茶叶中溴氰菊酯残留进行分析，测得检测限为1．35×10-2pg／ml，加标回收率为90％-125％。结论该单抗可用于农产品中溴氰菊酯类残留ELISA检测方法的建立。

Objective To prepare the monoclonal antibody （McAb） against dehamethrin （DTM）. Methods Schiff base meth- od was used to synthesize dehamethrin antigen which was used to immunize BALB/e mice. The hybridoma lines that secreted the monoclonal antibody （McAb） against deltamethrin were screened by fusing splenocytes and SP2/0 myelona cells and the an- tibodies were characterized by ELISA and SDS - PAGE. Results It showed that the antibodies were lgG1 subtype, the molecu- lar weight was 168 kDa, the affinity constant of McAb with coated complete antigen was 4.75 × 108 L/mol and the antibody was specific. Deltamethrin in tea was analyzed by ELISA. The limit of detection was 1.35 × 102 pg/ml and the recovery was 90% - 125%. Conclusion The MeAb can be used to establish the ELISA method for detection of DTM in agricultural products.