Preparation And Characterisation Of Monoclonal Antibodies Against Deoxynivalenol

ITALIAN JOURNAL OF ANIMAL SCIENCE(2020)

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Abstract
To prepare broad-spectrum and specific monoclonal antibodies (mAb) against deoxynivalenol (DON), DON-BSA was synthesised by the carbonyl diimidazole method as an artificial antigen to immunise Balb/C mice, and the coated DON-OVA was synthesised by the carbodiimide method to detect anti-DON antibodies. Monoclonal antibodies were screened by indirect ELISA and indirect competitive ELISA. A hybridoma cell line (4F3) capable of stably secreting DON antibody was obtained. The titre of antibody in the culture supernatant was 1:1.28 x 10(3), and the titre of the ascites antibody was 1:3.2 x 10(5). The monoclonal antibody homology was IgG1/lambda. The half inhibitory concentration (IC50) of DON was 9.84 ng/mL. The cross-reaction rates with 3-Ac-DON and 15-Ac-DON were 60.44% and 52.04%, respectively, but no cross-reaction with other mycotoxins was observed. The results showed that the anti-DON monoclonal antibodies prepared in this experiment could recognise not only DON but also 3-Ac-DON and 15-Ac-DON, which could provide materials for the next step to establish a method to detect DON and its similar compounds.
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Key words
Anti-deoxynivalenol, artificial antigen, monoclonal antibody, characterisation analysis
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