抗阿莫西林多克隆抗体的制备

目的]为研究阿莫西林残留的免疫学检测方法奠定基础。方法]采用N-羟基琥珀酰亚胺活性酯(NHS)法将阿莫西林分别与牛血清白蛋白(BSA)、卵清蛋白(OVA)偶联,制备免疫抗原AMX-BSA和检测抗原AMX-OVA,用AMX-BSA免疫成年兔以获得高效价的多克隆抗体。结果]免疫抗原AMX-BSA紫外光谱具有药物和蛋白的叠加特征,其最大吸收峰在276 nm处,说明载体蛋白BSA与AMX成功偶联,可用于动物免疫。检测抗原AMX-OVA最大吸收峰在275 nm处,说明载体蛋白OVA与AMX成功偶联,可用作检测抗原进行包被。间接ELISA检测结果表明,2只兔抗血清效价较高,均达1∶32 000以上,说明有特异性抗体产生。结论]该研究成功制备了抗阿莫西林多克隆抗体,也进一步证明药物偶联成功。

Preparation of Polyclonal Antibody against Amoxicillin

Objective] The research aimed to lay the foundation for studying the immunological method for detecting the residue of amoxicillin.Method] Amoxicillin were coupled with bovine serum albumin(BSA)and ovalbumin(OVA)by using N-hydroxysuccinimide actived ester method to prepare the immune antigen AMX-BSA and detection antigen AMX-OVA.The adult rabbits were immunized with AMX-BSA to obtain the polyclonal antibody with high titer.Result] The ultraviolet spectrum of immune antigen AMX-BSA had the stacking characteristics of medicament and protein and its maximum absorption peak was 276 nm,which indicated that vector protein BSA was successfully coupled with AMX and it could be used in animal immunization.The maximum absorption peak of detection antigen AMX-OVA was 275 nm,indicating vector protein OVA was successfully coupled with AMX and it could be taken as detection antigen for coating.The results of indirect ELISA detection showed that the antiserum titers of 2 rabbits were higher and both reached over 1∶32 000,which indicated that the specific antibody was produced.Conclusion] In this research,the polyclonal antibody against amoxicillin was successfully prepared,which further proved that the drugs were successfully coupled.