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

• Published in: Journal of agricultural and food chemistry 1993-02, Vol.41 (2), p.329-333
• Main Authors: Huang, Xuan, Chu, Fun Sun
• Format: Article
• Language: English
• Subjects: Analysis of the immune response. Humoral and cellular immunity; Antibody production; ANTICORPS; ANTICORPS MONOCLONAL; ANTICUERPOS; ANTICUERPOS MONOCLONALES; Biological and medical sciences; ELISA; Fundamental and applied biological sciences. Psychology; Fundamental immunology; HIBRIDOMA; HYBRIDOME; Immunobiology; MICOTOXINAS; MYCOTOXINE; REACCION ANTIGENO-ANTICUERPO; REACTION ANTIGENE ANTICORPS; TEST ELISA
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf00026a038

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