甲氰菊酯多克隆抗体的制备

Preparation of Polyclonal Antibody for Synthetic Pyrithroid Pesticide Fenpropathrin

利用水溶性碳二亚胺法将甲氰菊酯的合成前体甲氰菊酸与牛血清蛋白偶联,合成了甲氰菊酯的免疫半抗原,免疫新西兰大白兔获得了甲氰菊酯多克隆抗体。通过紫外扫描分析,免疫原中甲氰菊酸分子与蛋白质分子的偶联比为8.8:1,50%饱和硫酸铵法纯化抗体,间接ELISA法测定多克隆抗体效价达1:12800,最适甲氰菊酸-OVA包被抗原质量浓度为1.0mg/L;最低检出限为8.5μg/L。以抗体与甲氰菊酯的反应率为100%,抗体与其他4种菊酯类农药交叉反应率很低,与溴氰菊酯交叉反应较高。结果表明:制备的甲氰菊酯多克隆抗体具有较高的特异性,可用于甲氰菊酯残留的检测。

A polyclonal antibody specific for fenpropathrin was prepared. Hapten fenpropathrin acid was coupled with carrier protein bovine serum albumin to form an antigen FA-BSA via 1-ethyl-3-（3-dimethyl aminopropyl） and carbodiimide hydrochloride. The conjugates used to immunize New Zealand white rabbits were identified with UV scanning spectrum and the couple rate of the antigen was 8.8：1. After the antisera were obtained and purified using 50% saturated ammonium sulfate method, the specificity of PAb was detected by indirect ELISA （enzyme-linked immunosorbent assay）. It showed that the optimal concentration of coating antigen and PAb against fenpropathrin prove to be 1.0 mg/L and 1：12800, respectively. The limited detectable concentration was 8.5 μg/L. The cross-reaction rate between PAb and other pyrethroid pesticides was low, except deltamethrin. The results showed that the anti-fenpropathrin PAb had a high specificity and was good enough for immunoassay of fenpropahtrin.