Synthesis and identification of parathion complete antigen and its immune effect
Parathion is a colorless and odorless liquid. It is a broad-spectrum insecticide with the functions of contact, stomach poisoning and fumigation. However, if people inhale or ingest, it will cause poisoning and serious death. Recently, researchers from the School of Food Science and Technology of Huazhong Agricultural University published a paper aimed at synthesizing artificial antigens with strong immunogenicity, laying a foundation for the preparation of monoclonal antibodies against parathion and rapid establishment of methods for detecting pesticide residues in foods. The study indicated that the successful synthesis of two parathion-complete antigens was better than that of the PA-1-BSA group in the immunized Balb/c mice. The article was published in the No. 01 issue of the Journal of Preventive Medicine of the People's Liberation Army in 2013.
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Two methods were used to synthesize carboxylated parathion: one with parathion as raw material to modify one end of nitro group; the other with trichlorosulfate as raw material to transform one end of phosphorus-sulfur bond. The EDC method was coupled with bovine serum albumin (BSA) to prepare immunogens (PA-1-BSA and PA-2-BSA) and to immunize Balb/c mice, respectively. The antiserum titer was determined by indirect ELISA, the sensitivity of the assay was determined by competition ELISA, and the immune effect was compared.
SDS-PAGE electrophoresis showed that the two complete antigens were successfully synthesized. The coupling ratios of the immunogens identified by mass spectrometry were: N(PA-1/BSA)=7.2, N(PA-2/BSA)=15.4; mice The indirect ELISA showed that the titer of PA-2-BSA group was significantly higher than that of PA-1-BSA group. The indirect competitive ELISA showed that the half-inhibitory concentration (IC50) of PA-2-BSA group was 1μg/ml. Better than PA-1-BSA group (IC50 >100 μg/ml).