Production and characterization of monoclonal and polyclonal antibodies against the mycotoxin cyclopiazonic acid

Polyclonal antibodies (Pabs) against cyclopiazonic acid (CPA) were obtained from both mice and rabbits after they were immunized with CPA-bovine serum albumin (BSA) or ethylenediamine-modified BSA. An indirect enzyme-linked immunosorbent assay (ELISA) was developed for antibody titer determination and for CPA assay. Antibody titer reached 9000 9 weeks after immunization. Competitive ELISA revealed that the concentrations causing 50% inhibition binding (ID50) of the rabbit Pabs to the solid-phase CPA-keyhole limpet hemocyanin by CPA and CPA-imine were 62 and 3388 ng/mL, respectively. Four hybridoma cell lines (5C8B2, 5C8D1, 5C8D3, 5C8F3) that produced IgG1 and kappa-chain isotype monoclonal antibodies (Mabs) with a binding constant in the range 6.1 X 10(8) to 2.0 X 10(10) L/mol for CPA were generated. The ID50 of the Mab elicited by these clones ranged from 0.014 to 0.17 ng/mL for CPA and from 3.34 to 44.2 ng/mL of CPA-imine, respectively. The linear portion of the standard curve for the Mab-based ELISA (Mab from clone 5C8B2) for CPA was in the range 0.001-0.5 pg/assay. Neither Mabs nor Pabs cross-reacted with tenuazonic acid, D-lysergic acid, lysergol, and tryptophan