Generation and characterization of a monoclonal antibody to penicillic acid from Penicillium cyclopium

Abstract

Penicillic acid is one of the main mycotoxins in moldy feedstuff and has toxic effect on livestock and poultry and probably humans due to food chain transmission. The objective of this study was to generate and characterize a monoclonal antibody to penicillic acid for the efficient detection of penicillic acid from Penicillium cyclopium by immunological methods. To this end, penicillic acid was conjugated to bovine serum album (BSA) using the Mannich reaction and coupled with ovalbumin
(OVA) by the method of 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride (EDC) to generate artificial antigens penicillic acid-BSA and penicillic acid-OVA. A hybridoma cell line was obtained after fusion of mouse SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with artificial antigen penicillic acid-BSA. A monoclonal antibody specific against penicillic acid was produced in vivo by this hybridoma cell line. Further analysis revealed that the monoclonal antibody to penicillic acid was of the IgG1 subtype, with a titer of 1: 2.05 × 105. The antibody to penicillic acid had no or less cross-reaction with mycotoxins, including aflatoxin B1, zearalenone, T-2 toxin and fumonisins and more importantly, it assumed an affinity of about 1.54 × 108 liters per mol. Our ability to produce a monoclonal antibody to penicillic acid provides necessary groundwork for the effective detection of penicillic acid in various tissues of animals and human, using the immunocytochemistry,
Western blots and enzyme-linked immunosorbent assay (ELISA).