利用基因工程抗体建立的新型双抗夹心ELISA法检测Cry1Ac毒素

Establishment of a new double-antibody sandwich ELISA method for detection of Cry1Ac toxin with genetically engineered antibody

为明确新型双抗夹心ELISA方法检测Cry1Ac毒素的效果,分别用纯化后的抗Cry1Ac单链抗体和anti-Cry1Ac兔多克隆血清(Anti-Cry1Ac-PAbs)作为捕获抗体和检测抗体,通过方阵滴定确定其最佳工作浓度,建立双抗夹心ELISA法。再用优化后的双抗夹心ELISA方法对检测Cry1Ac毒素的敏感性、特异性和回收率进行鉴定。结果表明,新型双抗夹心ELISA法最低检测限为0.91 ng/ml,线性检测范围为1.04 ng/ml^3.49μg/ml。5种稻米中Cry1Ac毒素的平均回收率为69.42%~87.95%,变异系数为1.19%~6.50%。

The purified anti-Cry1Ac single chain antibody was used as capture antibody, and genetically engineered anti-Cry1Ac-PAbs was used as detection antibody to develop a double-antibody sandwich ELISA for detection of Cry1Ac toxin. The results showed that the limit of detection of Cry1Ac toxin was 0. 91 ng/ml, with the linear range from 1. 04 to 3. 49 ng/ml. The average recovery of Cry1Ac toxin in five kinds of rice was 69. 42%-87. 95%, with coeffi-cient of variation of 1. 19%-6. 50%.