兔抗IVM多克隆抗体的制备

The Preparation of Polyclone Antibody for Indirect Competitive ELISA of Ivermectin Residue

将伊维菌素(IVM)进行化学修饰,引入羧基活性基团,合成具有半抗原结构特征的伊维菌素半琥珀酸酯(IVM-HS);然后采用NHS法和混合酸酐法将半抗原与聚-L-赖氨酸(PLL)和卵清蛋白(OVA)相偶联,制备人工免疫原(PLL-IVM-HS)和包被原(OVA-IVM-HS);经凝胶电泳鉴定,用人工免疫原免疫新西兰大白兔制备多克隆抗体(pAb),间接ELISA测定其效价。结果人工免疫原偶联成功,分子结合比为25.3∶1,获得了高价、敏感、特异性的IVMpAb,为IVM残留免疫学检测法的建立奠定了基础。

The active group carboxyl was introduced to Ivermectin （IVM） and formed IVM half succinaate （IVM-HS） which had hapten structure by chemical modification. The method of NHS was used to conjugate IVM-HS to PLL and obtained artificial immunogen PLL-IVM-HS, the coating antigen OVA-IVM-HS was obtained by mixed anhydride reaction method. SDS-PAGE was used to identify PLL-IVM-HS. New zealand white rabbits were immunized with PLL-IVM-HS, the titre of polyclonal antibody （pAb） was detected by indirect ELISA. The results showed that PLL-IVM-HS had been synthesized successfully and its conjugation ratio of IVM-HS to PLL was about 25.3：1, the high-titer, sensitive and specific IVM pAb had been produced in sera of immunized new zealand white rabbits. This made it possible to establish immunoassay of IVM residues in feed and animal food.