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

作　　者：张留圈 张霄 刘媛[2,3,4] 徐重新 张存政[2,3,4] 谢雅晶 颜春荣[5] 寇莉萍 刘贤金[2,3,4] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100 [2]江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地 [3]农业部农产品质量安全控制技术与标准重点实验室 [4]江苏省农业科学院食品质量安全与检测研究所,江苏南京210014 [5]江苏省产品质量监督检验研究院,江苏南京210007

出　　处：《江苏农业学报》2014年第3期634-639,共6页Jiangsu Journal of Agricultural Sciences

基　　金："973"项目(2012CB722505);国家自然科学基金项目(31201535);江苏省农业自主创新基金项目[CX(12)5042];江苏省质量技术监督局科技项目(KJ112505)

摘　　要：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%.

关 键 词：基因工程抗体 Cry1Ac毒素 

分 类 号：TS207.53[轻工技术与工程—食品科学]