溴氰菊酯人工抗原及多克隆抗体制备

Preparation of Deltamethrin Artificial Antigen and Its Polyclonal Antibody

以二溴菊酸和4-[(4-硝基苯)氧基]苯甲醛为原料经4步反应合成了溴氰菊酯半抗原1-氰基[4-(4-氨基苯)苯基]甲基-2,2-二甲基3-(2',2'-二溴乙烯基)环丙烷羧酸酯;用重氮化法将溴氰菊酯半抗原分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)相偶联制备免疫抗原和包被抗原,紫外吸收法估算偶联比分别为11∶1和8∶1;以BSA偶联物作免疫抗原免疫日本大耳白兔制备溴氰菊酯多克隆抗体,间接非竞争ELISA法测定两个抗血清效价分别为100000和80000,间接竞争ELISA法测定抗体特异性结果表明所制备的抗体测定溴氰菊酯的IC50值为0.16 mg/L,与其他供试农药的交叉反应率均≤1%。制备的溴氰菊酯人工抗原和多克隆抗体为研究建立溴氰菊酯农残免疫快速测定方法奠定了基础。

The hapten of deltamethrin, （3-（3-aminophenoxy）phenyl）（cyano）methyl 3-（2,2-dibromovinyl）-2,2- dimethylcyclopropanecarboxylate, was synthesized. The structure of which was identified by 1H NMR. Hapten was conjugated to bovine serum albumin （BSA） and ovalbumin （OVA） by diazotization method, two artificial antigens were used as antigen and coating antigen. Antigens were confirmed by ultraviolet spectrum. The coupling ratio were 11：1 and 8：1. Two rabbits were immunized, the antiserum was examined by indirect ELISA and indirect competitive ELISA experiments, the titer of the two rabbits was 100000 and 80000 separately. Results of a competitive indirect ELISA showed that the ICs0 value of deltamethrin was 0.16 mg/L, and that there was less than 1% cross-reactivity with other tested pesticides. Therefore, the artificial antigen and its polyclonal antibody paved way for development of fast immunoassay methods for deltamethrin residue.