Preparation of Bispecific Antibodies Against Zearalenone and Deoxynivalenol and Construction of Detection Methods

Abstract:

Zearalenone (ZEN) and deoxynivalenol (DON) are common mycotoxins found in grain, and long-term dietary exposure can cause serious harm to organisms. Currently, simultaneous immunoassays for ZEN and DON mostly rely on their respective monoclonal antibodies. However, the preparation of traditional monoclonal antibodies requires long production cycles and high costs. In this study, bispecific antibodies(Bis-scFv) that could simultaneously recognize both ZEN and DON were obtained in a short time using recombinant antibody expression technology. An indirect competitive enzyme-linked immunosorbent assay(IC-ELISA) based on Bis-scFv was established. The half-maximal inhibitory concentrations(IC₅₀) for ZEN and DON standard curves was determined to be 20.64 ng/mL and 132.29 ng/mL, respectively. Bis-scFv exhibited excellent specificity with no significant cross-reactivity with other mycotoxins. Moreover, IC-ELISA was used to conduct spike recovery experiments for ZEN and DON in corn, yielding recovery rates ranging from 86.02% to 108.14%. The study demonstrates that the developed Bis-scFv has the potential for future application in the development of synchronous and rapid detection of ZEN and DON in grain samples.