Preparation of Artificial Antigen and ELISA Identification of Rabbit Polyclonal Antiserum for the Detection of Glyphosate

doi:10.7506/spkx1002-6630-201704011

Abstract

Abstract: To obtain polyclonal antiserum with high sensibility and specificity, artificial antigens were synthesized by coupling glyphosate (PMG) with the carrier proteins bovine serum albumin (BSA) and ovalbumin (OVA) separately by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and glutaraldehyde (GA) methods in this study. As a result, the immune antigen PMG-BSA and detection antigen PMG-OVA were obtained and identified by UV spectroscopy and SDS-PAGE. New Zealand white rabbits were immunized with PMG-EDC-BSA and PMG-GA-BSA, respectively to obtain polyclonal antisera. The titer of the antisera was determined by indirect ELISA and their sensibility and specificity were determined by indirect competitive ELISA. The results showed that the titer of antiserum of all three immunized rabbits was higher than 1:104. The antiserum of rabbit 2 exhibited the highest sensibility with an IC50 of 30.9 μg/mL and good specificity. The successful synthesis of artificial antigens and the obtainment of PMG polyclonal antiserum with high sensibility and specificity may lay the foundation for rapid immunological detection of PMG.

Key words: glyphosate, artificial antigen, polyclonal antiserum, ELISA

CLC Number:

S482.4

WANG Jing, WANG Yao, WANG Fangyu, DENG Ruiguang, HU Xiaofei, YU Qiuying, HAO Junfang, XING Yunrui, HOU Yuze. Preparation of Artificial Antigen and ELISA Identification of Rabbit Polyclonal Antiserum for the Detection of Glyphosate[J]. FOOD SCIENCE, 2017, 38(4): 65-69.

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Preparation of Artificial Antigen and ELISA Identification of Rabbit Polyclonal Antiserum for the Detection of Glyphosate

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