Production and characterization of a monoclonal anti-anti-idiotype antibody against fumonisin B-1

A monoclonal anti-anti-idiotype antibody (mAb3) against fumonisin B-1 (FmB1 ) was produced from the hybridoma cell line 7C7F4, which was generated by t he fusion of P3/NS-1/1-AG4-1 myeloma cells with spleen cells isolated from a Balb/c mouse that had been immunized with the Fab fragments of affinity-p urified anti-idiotype antibodies. The mAb3 belongs to the immunoglobulin M, kappa light chain. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were established f or antibody characterization and toxin analysis. In an idc-ELISA using FmB1 -ovalbumin (OVA) as the coating antigen, the concentrations causing 50% inh ibition of binding (IC50) of mAb3 to the solid-phase FmB1-OVA by free FmB1, FmB2, and FmB3 were found to be 75, 95, and 450 ng/mL, respectively. In th e dc-ELISA, the concentration causing IC50 of FmB1-horseradish peroxidase t o the solid-phase mAb3 by free FmB1 was found to be 233 ng/mL. Analysis of 12 samples naturally contaminated with fumonisins with mAb3-based idc-ELISA and polyclonal-based dc-ELISA showed a good correlation between these two methods with a correlation coefficient of 0.76 at p < 0.02. The linear regr ession slope was found to be y[polyclonal ELISA] = 0.87x[mAb3 ELISA] - 52 p pb.