Liu Yuanyuan, Jiang Dongjian, Lu Xin, Wang Wei, Xu Yang, He Qinghua
State Key Laboratory of Food Science and Technology, Sino-German Joint Research Institute, Nanchang University , No. 235 Nanjing East Road, Nanchang 330047, China.
J Agric Food Chem. 2016 Oct 19;64(41):7882-7889. doi: 10.1021/acs.jafc.6b02978. Epub 2016 Oct 11.
The widespread use of Cry proteins in transgenic plants for insect control has raised concerns about the environment and food safety in the public. An effective detection method for introduced Cry proteins is of significance for environmental risk assessment and product quality control. This paper describes a novel phage mediated immuno-PCR (iPCR) for the ultrasensitive determination of Cry proteins based on nanobodies. Three nanobodies against Cry1Ac protein were obtained from a naı̈ve phage displayed nanobody library without animal immunization process and were applied to the iPCR assay for Cry1Ac. The phage-mediated iPCR for Cry1Ac based on nanobodies showed a dynamic range of 0.001-100 ng/mL and a limit detection of 0.1 pg/mL. Specific measurement of this established method was performed by testing cross-reativity of other Cry1Ac analogues, and the result showed negligible cross-reactivity with other test Cry proteins (Cry1Ab, Cry1F, Cry3B). Furthermore, the phage-mediated iPCR based on nanobody should be easily applicable to the detection of many other Cry proteins.
转基因植物中广泛使用Cry蛋白来控制昆虫，这引发了公众对环境和食品安全的担忧。一种有效的导入Cry蛋白检测方法对于环境风险评估和产品质量控制具有重要意义。本文描述了一种基于纳米抗体的新型噬菌体介导免疫PCR（iPCR）用于超灵敏测定Cry蛋白。从未经动物免疫过程的天然噬菌体展示纳米抗体文库中获得了三种针对Cry1Ac蛋白的纳米抗体，并将其应用于Cry1Ac的iPCR检测。基于纳米抗体的Cry1Ac噬菌体介导iPCR的动态范围为0.001 - 100 ng/mL，检测限为0.1 pg/mL。通过测试其他Cry1Ac类似物的交叉反应性对该既定方法进行特异性测量，结果显示与其他测试Cry蛋白（Cry1Ab、Cry1F、Cry3B）的交叉反应可忽略不计。此外，基于纳米抗体的噬菌体介导iPCR应该很容易应用于许多其他Cry蛋白的检测。
