黄曲霉毒素B_1标准替代物的制备及其在花生样品检测中的应用

Preparation and application of surrogate calibrator for detecting aflatoxin B_1 in peanut

以抗黄曲霉毒素B1(AFB1)单克隆抗体的F(ab')2片段为抗原免疫兔子,得到AFB1抗独特型抗体。经过酶联免疫吸附法(ELISA)条件的优化,建立AFB1抗独特型抗体最佳竞争抑制曲线。该曲线与AFB1竞争抑制曲线相比较可知,AFB1抗独特型抗体与AFB1之间存在指数增长关系,且相关性系数R为0.999 8。以AFB1抗独特型抗体浓度与其相应的抑制率作标准曲线,ELISA法测定花生中AFB1添加回收率,范围在90.4%～100.2%之间。综上,AFB1抗独特型抗体可以作为AFB1无毒替代标准品。

Anti-idiotype antibodies,used as surrogate of AFB1,were generated by immunizing the rabbit with F（ab′）2 fragments of anti-AFM1 mAb.After optimizing physicochemical factors of ELISA,a competitive inhibition curve of anti-idiotype antibodies was obtained.An excellent exponential growth correlation（R=0.999 8） was observed between AFB1 standard solutions and anti-idiotype antibody solutions.The recovery of AFB1 ranged from 90.4% to 100.2% in peanuts.Therefore,the anti-idiotype antibodies can be used for detecting AFB1 in peanuts as surrogate calibrators.