[Preparation of anti-zearalenone monoclonal antibody and preliminary establishment of colloidal gold immunochromatographic assay for zearalenone].

Objective: To prepare the monoclonal antibody against zearalenone (ZEN) toxin and preliminarily establish the colloidal gold immunochromatographic detection method for ZEN.

Methods: The artificial antigen ZEN-BSA and ZEN-OVA were prepared by active ester method. Mice were immunized with ZEN-OVA and monoclonal antibodies against ZEN were prepared by regular cell fusion and subcloning approach. The titer, subtype and specificity of the antibodies were identified by ELISA. To establish a method of colloidal gold immunochromatographic assay (GICA) for the determination of ZEN, colloidal gold binding cushion was coated with anti-ZEN monoclonal antibody-colloidal gold complex, and the synthetic ZEN-BSA and goat anti-mouse immunoglobulins were sprayed on cellulose nitrate film to form the test (T) band and control (C) band.

Results: Identified by SDS-PAGE and UV spectroscopy, the artificial antigen ZEN-BSA and ZEN-OVA were successfully coupled respectively. One hybridoma line (1G4) which could secrete monoclonal antibody specifically against ZEN was obtained by three-time cloning. The ascites titer of this monoclonal antibody reached 1:1.6×10(5);. The antibody subtype was IgG2b. The IC50; to ZEN was 10.2 ng/mL, and the detction limit was 0.58 ng/mL. In addition, the mAb was proved highly specific for ZEN. ZEN metabolites α-zearalanol, β-zearalanol, zearalanone and other similar toxins deoxynivalenol, fumonisin B1, ochratoxin A showed very low or no cross-activity with the monoclonal antibody. The whole assay using the prepared colloidal gold immunochromatographic strip could be finished in 5 min, which could be read by naked eyes. The limit of detection was 100 ng/mL.

Conclusion: The anti-ZEN mAb was successfully prepared. And the gold immunochromatographic assay(GICA) for ZEN was preliminarily established with the limit of detection being 100 ng/mL.