Preparation Of Monoclonal Antibodies Against Fumonisin B₁ And Development Of ELISA Kit

Fumonisin B1 (FB1) is one of mycotoxins produced by Fusarium moniliforme which has pulmonary toxicity, neurotoxicity, immune system toxicity, carcinogenicity and many kinds other toxicity. Pollution range of FB1 is very wide, the main pollution is maize and its products, also can pollute the wheat, rice and other food and its (?) to human and animal, thus it becomes more concerned by the people in food security. The International Center for research on cancer (IARC) have FB] identified as a class 2B carcinogen (make it possible for human induced cancer). At present, the detection of FB1 is mainly based on the measurement, because the equipment is expensive and complicated operation, thus there are certain limitations. Immunological detection method which based on monoclonal antibody not only overcomes the above problems, but also has higher sensitivity, and more favorable for popularization.This study used 4G5 cells which preserved in our laboratory and could secreted against FB1 monoclonal antibodies, for further screening. The cell line 4G5-A4 which we screened could stably secrete monoclonal antibody and had higher titers. Then, we used this cell line for preparating and purifing anti-FB1 monoclonal antibody. The protein concentration of unpurified antibody was 12.58 mg/mL, purified antibody was 1.62 mg/mL, and the recovery rate of IgG was 25.63%. The subtype of antibody was IgG2b, and the affinity was 1.28×108 L/mol. Monoclonal antibody had no cross reaction with AFB1, CIT, CTX, DON, but a slight cross-reactivity with T2. The obtainment of high quality monoclonal antibody, laied a foundation for further study of FB1 immune detection technology.Used the anti-FB1 monoclonal antibodies, we established indirect competitive ELISA method, and optimized the parameters. The concentration of coating antigen was 0.05 ug/mL, and the dilution of antibody was 1:4000. The detection linear range of FB1 detection method was 1.29-804.38 ng/mL, and the lowest detection limit was 0.27 ng/mL. Average recovery rate of mock sample was 96.561%～109.025%, and the coefficient of variation was less than 10%. The average concentration of FB1 detected in real samples was 195.80 ng/mL, and the maximum concentration was 355.95 ng/mL. To a certain extent, The detection method could be applied to safety testing and risk assessment of corn and corn products.