Citation: TAN Gui-Yu, ZHU Yi-Fan, ZHANG Man, HE Qing-Qing, CHEN Xiao-Jiao, ZHANG Kun, WANG Bao-Min. Development and Application of Time-Resolved Fluorescence Lateral Flow Immunoassay for Bt Cry1Ab/Ac Protein Detection in Transgenic Plant[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(5): 752-758. doi: 10.19756/j.issn.0253-3820.191515 shu

Development and Application of Time-Resolved Fluorescence Lateral Flow Immunoassay for Bt Cry1Ab/Ac Protein Detection in Transgenic Plant

TAN Gui-Yu ^1,2 ,
ZHU Yi-Fan ¹ ,
ZHANG Man ¹ ,
HE Qing-Qing ¹ ,
CHEN Xiao-Jiao ^1,, ,
ZHANG Kun ^2,3 ,
WANG Bao-Min ^1,,

1.
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China;
2.
Guangxi Botanical Garden of Medicine Plant, Nanning 530023, China;
3.
Fujian Antiorigin Biotechnology Co, Ltd., Fuzhou 350026, China

Corresponding author: CHEN Xiao-Jiao, WANG Bao-Min,
Received Date: 25 August 2019
Revised Date: 1 April 2021

Fund Project: Supported by the National Key Research and Development Program of China (No. 2018YFC1602900) and the Fuzhou Cangshan District Science and Technology Plan Project (No. 2017C03).

By now, genetically modified crops have been widely planted in the world. It is an urgent need not only to conduct qualitative testing of transgenes crop, but also to conduct rapid quantitative analysis of exogenous protein content expressed in transgenic crops, so as to judge the resistance to insects or herbicides. Two mouse monoclonal antibodies that recognize different epitopes of Bt Cry1Ab/Ac have been prepared in the previous work. On this basis, time-resolved fluorescent strips of Bt Cry1Ab/Ac were prepared using fluorescent microspheres as markers in this work. Combined with portable fluorescence immunoanalyzer, a lateral flow immunoassay was developed for the quantitative analysis of exogenous protein Bt Cry1Ac in transgenic crops. The detection time was 20 min, the linear equation of the standard curve was y=0.5804x-1.9123, with correlation coefficient (R²) of 0.998. The limit of quantitation was 1.5 ng/mL. With the established method, the content of Bt Cry1Ab/Ac protein in samples was determined, and the results were compared with those of commercial kit with detection time of 1.5 h. The results of these two methods correlated well (R²=0.999). The time-resolved fluorescent strip established here could be used as a rapid and accurate quantitative method for determination of Bt cry1Ab/Ac in transgenic crops.
- Transgenic crop,
- Bt Cry1Ab/Ac,
- Time-resolved fluorescent lateral flow immunoassay
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