Phage Medicated Immuno-PCR for Ultrasensitive Detection of Cry1Ac protein based on Nanobody.

Journal of agricultural and food chemistry

PubMedID: 27684201

Liu Y, Jiang D, Lu X, Wang W, Xu Y, He Q. Phage Medicated Immuno-PCR for Ultrasensitive Detection of Cry1Ac protein based on Nanobody. J Agric Food Chem. 2016;.
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. In this report, we describe a novel phage medicated immuno-PCR (iPCR) for ultrasensitive determination of Cry proteins based on nanobodies. Three nanobodies against Cry1Ac protein were obtained from a naïve phage displayed nanobody library without animal immunization process and were applied to iPCR assay for Cry1Ac. The phage medicated immuno-PCR 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, the result showed negligible cross-reactivity with other test Cry proteins (Cry1Ab, Cry1F, Cry3B). Furthermoer, the phage medicated iPCR based on nanobody should be easily applicable to detect many other Cry proteins.