Phage-displayed nanobody based double antibody sandwich chemiluminescent immunoassay for the detection of Cry2A toxin in cereals
文献类型： 外文期刊
作者： Qiu, Yulou 1 ; Li, Pan 1 ; Liu, Beibei 1 ; Liu, Yuan 1 ; Wang, Yulong 1 ; Tao, Tingting 1 ; Xu, Junli 1 ; Hammock, Bruce 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Key Lab Control Technol & Stand Agroprod Safety &, State Key Lab Cultivat Base,Minist Sci & Technol, Minist Agr,Inst Food Safety & Nutr,Lab Food Qual, Nanjing 210014, Jiangsu, Peoples R China
2.China Jiliang Univ, Zhejiang Proceincial Key Lab Biometrol & Inspect, Hangzhou 310018, Zhejiang, Peoples R China
3.Univ Calif Davis, Dept Entomol, Lab Pesticide & Nematol Biotechnol, Davis, CA 95616 USA
4.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Jiangsu, Peoples R China
关键词： Cry2A toxin; phage-displayed nanobody; chemiluminescence; sandwich immunoassay
期刊名称：FOOD AND AGRICULTURAL IMMUNOLOGY （ 影响因子：3.101； 五年影响因子：2.573 ）
ISSN： 0954-0105
年卷期： 2019 年 30 卷 1 期
页码：
收录情况： SCI
摘要： In this study, we developed a double antibody sandwich chemiluminescent immunoassay (DAS-CLIA) based on phage-displayed nanobodies for the determination of Cry2A toxin. Nine phage-displayed nanobodies specifically to Cry2A were obtained from a naive nanobody library after four rounds of biopanning. The phage-displayed nanobody P2 with high affinity binding was selected as the detection antibody for the sensitive DAS-CLIA development. The proposed method exhibited high sensitivity with a limit of detection of 0.09 ng/mL, has a wide linear range for Cry2A toxin detection, in the range of 0.1-1000 ng/mL and negligible cross-reactivities to the other Bt toxins. The recoveries of Cry2A toxin spiked in cereal samples (rice, wheat, and corn) ranged from 82.7% to 118%, with a coefficient of variation of less than 10%. This study demonstrated that the DAS-CLIA procedure based on phage-displayed nanobodies may provide an alternative sensitive method for the detection of Bt toxins in agri-products. [GRAPHICS] .
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