绿色无毒黄曲霉毒素B_1免疫检测方法

Green and Nontoxic Enzyme-Linked Immunosorbent Assay for Aflatoxin B1 Detection

摘要:

建立了一种基于抗独特型抗体的绿色无毒的ELISA方法,并用于检测面粉中黄曲霉毒素B1。通过酶解2种抗黄曲霉毒素小鼠单克隆抗体(11A9和1G3)制备F(ab’)2片段,并将其免疫新西兰大白兔制备抗独特型抗体。最终得到两种抗独特型抗体,并对抗独特型抗体和AFB1进行相关性分析。通过实际样品添加回收,其批内回收率为115.60%~121.88%,变异系数在5%以内;而批间回收率为111.89%~126.98%,变异系数低于8%。分析结果准确可靠,表明此无毒绿色ELISA是一种安全可靠的AFB1分析方法。

Abstract:

A environmental-friendly and nontoxic enzyme-linked immunosorbent assay(ELISA) for the detection of aftoxin B1(AFB1) in wheat flour sample was developed. In this method,the standard substance was substituted by the anti-idiotype antibody(anti-id antibody). The anti-id antibodies were prepared by immunizing the rabbits with the F(ab’)2 fragments that were obtained by digesting the monoclonal antibodies(11A9 and 1G3) against AFB1. Two kinds of anti-id antibodies was prepared and both could mimic the three-dimensional structural of AFB1 and caused specific inhibition to the 11A9 mAb. The correlation of the AFB1 and anti-id antibodies were analyzed by regression. The green ELISA based on anti-id antibody was applied in the analysis of spiked wheat flour sample. The recovery intra-and inter-assay was between 111.89% and 126.98%,respectively,while the CV less than 10%. The results indicated that the environmentalfriendly and nontoxic ELISA could be used for actual samples.