Establishment of an ELISA Method for Quantitative Detection of PAT/pat in GM Crops

Weixiao Liu (), Lixia Meng, Xuri Liu, Chao Liu and Wujun Jin ()
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Weixiao Liu: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Lixia Meng: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Xuri Liu: Department of Food and Biological Engineering, Handan Polytechnic College, Handan 056001, China
Chao Liu: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
Wujun Jin: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Agriculture, 2022, vol. 12, issue 9, 1-11

Abstract: The phosphinothricin N-acetyltransferase gene ( pat ) is widely used to confer resistance to the herbicide phosphinothricin for genetically modified (GM) crops. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) is developed to detect PAT/pat in GM crops. Two anti-PAT/pat monoclonal antibodies (mAbs), 1F5-1F2 and 1B6-2D3, with titers of 1:1,024,000 and 1:896,000, respectively, against overexpressed His-PAT/pat, were screened out, raised, and characterized. An ELISA method was established with the 1F5-2F2 mAb for capture and the biotin-labeled 1B6-2D3 mAb for detection of PAT/pat. The linear detection range of the method was approximately 1.5625–12.5 ng/mL, with a sensitivity of 0.085 ng/mL and a coefficient of variation (CV) less than 5.0%. No cross-reactivity was found with other herbicide resistance proteins, especially PAT/bar. The established sensitive and specific ELISA was successfully applied in the detection of PAT/pat expression in GM crops.

Keywords: PAT/pat; quantitative detection; monoclonal antibody; ELISA; genetically modified crops (search for similar items in EconPapers)
JEL-codes: Q1 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 (search for similar items in EconPapers)
Date: 2022
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