Preparation of anti-zearalenone monoclonal antibodies and development of an indirect competitive ELISA for zearalenone

Abstract:

Zearalenone (ZEN) is a high toxic mycotoxin. How to detect ZEN from food and feed is important and challenging. Monoclonal antibody was prepared by means of immunization in mice with conjugates of ZEN and bovine serum albumin (BSA), and enzyme-linked immunosorbent assay for ZEN based on monoclonal antibodies was developed. Four hybridoma cell strains secreting anti-zearalenone monoclonal antibodies were cloned. The subclasses of the monoclonal antibody were characterized as Ig G1 for three and Ig G2b for one. One of the cell strains 2C9 was used to prepare ascites for its high bioactivity. The titer of purified ascites was 1/40 000. The indirect-competitive ELISA was developed with good specificity for ZEN, the half maximal inhibitory concentration (IC50) is 1.90 ng/mL, the detection limit is 0.051 ng/mL, and the detection range is 0.115?13.9 ng/mL. The recovery rate could get to 96.5%?113% while the sample containing ZEN 1.46?93.8 μg/kg. The method for detection was de-veloped, which could be applied to detect ZEN from a variety of grain and feed samples.

Key words:Zearalenone, Monoclonal antibody, Enzyme-linked immunosorbent assay, Recovery rate