Preparation and characterisation of monoclonal antibodies against deoxynivalenol

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 × 103, and the titre of the ascites antibody was 1:3.2 × 105. The monoclonal antibody homology was IgG1/λ. 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.Highlights This study demonstrated that an artificial antigen was synthesised by carbonyl diimidazole method, and a novel broad-spectrum, high affinity monoclonal antibody was developed. The anti-DON mAb developed in this study has better sensitivity, moreover the antibody was sensitive to DON and its derivatives. Therefore, it could be used for simultaneous monitoring of three mycotoxins. Results of this study will further lay the foundation for the establishment of immunological assays for the total amount of similar compounds.

Keywords

• anti-deoxynivalenol
• artificial antigen
• monoclonal antibody
• characterisation analysis