Phage-Mediated Immuno-PCR for Ultrasensitive Detection of CrylAc Protein Based on Nanobody
Phage-Mediated Immuno-PCR for Ultrasensitive Detection of CrylAc Protein Based on Nanobody
复制标题
基于纳米抗体的噬菌体介导免疫 PCR 超灵敏检测 CrylAc 蛋白
DOI：
10.1021/acs.jafc.6b02978
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发表时间：
2016-10-19
影响因子：
6.1
通讯作者：
He, Qinghua
中科院分区：
文献类型：
--
作者：
Liu, Yuanyuan;Jiang, Dongjian;He, Qinghua
Cry蛋白在转基因植物中的广泛使用引起了公众对环境和食品安全的担忧。建立一种有效的Cry蛋白检测方法，对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用于超灵敏测定Cry蛋白的新型噬菌体介导的免疫PCR（iPCR）。从未经动物免疫处理的天然噬菌体展示纳米抗体库中获得三个针对CrylAc蛋白的纳米抗体，并将其应用于CrylAc的iPCR测定。基于纳米抗体的噬菌体介导的CrylAc iPCR显示出0.001-100 ng/mL的动态范围和0.1 pg/mL的检测限。通过测试其他CrylAc类似物的交叉反应性来进行该建立的方法的具体测量，结果显示与其他测试Cry蛋白（CrylAb、Cry 1F、Cry 3B）的交叉反应性可忽略不计。此外，基于纳米抗体的噬菌体介导的iPCR应易于适用于许多其他Cry蛋白的检测。
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 CrylAc protein were obtained from a na ve phage displayed nanobody library without animal immunization process and were applied to the iPCR assay for CrylAc. The phage-mediated iPCR for CrylAc 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 CrylAc analogues, and the result showed negligible cross-reactivity with other test Cry proteins (CrylAb, Cry1F, Cry3B). Furthermore, the phage-mediated iPCR based on nanobody should be easily applicable to the detection of many other Cry proteins.