Production and characterization of monoclonal antibodies against fumonisin B1

Monoclonal antibodies (mAb) against fumonisin B1 (FmB1) were produced from a stable hybridoma cell line, 9C11E6, generated by the fusion of P3/NS-1/1-AG4-1 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with FmB1-keyhole limpet hemocyanin. The 9C11E6 mAb belong to the immunoglobulin GI (kappa chain) isotype with the highest specificity for FmB3. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were established for antibody characterization and toxin analysis. The concentrations causing 50% inhibition (IC(50)) of binding of FmB1- horseradish peroxidase with the antibodies by FmB1, FmB2, and FmB3 in the dc-ELISA were found to be 29.1, 17.5, and 4.1 ng ml(-1), respectively. The IC(50) values of the binding of 9C11E6 mAb to the solid-phase FmB1-ovalbumin by free FmB1, FmB2, and FmB3 in the idc-ELISA were found to be 20.1, 13.2, and 16.8 ng ml(-1), respectively. The average recovery of FmB1 (50-2000 ng g(-1)) added to cornmeal and subsequently extracted with phosphate-buffered saline in the dc-ELISA was found to be 71.3%. The mAb did not react with the hydrolyzed FmB1, sphingosine, sphinganine, or tricarballylic acid, but shouted weak cross-reactivity (about 6.8% of FmB1) with the Alternaria alternata f. sp. lycopersici toxin TA.