Development of a rapid magnetic bead-based immunoassay for sensitive detection of zearalenone

Here we demonstrate a novel magnetic bead-based enzyme-linked immunosorbent assay (MB-ELISA) for zearalenone (ZEN) detection. Firstly, an anti-ZEN monoclonal antibody (mAb) was prepared by hybridoma technique, and immobilized on carboxyl modified MBs to obtain mAb-MBs. In addition, the biotinylated ZEN-BSA was labelled by streptavidin-HRP for use as competitor. Based on the mAb-MBs and streptavidin-HRP labelled ZEN-BSA, a MB-ELISA which contains only one 20 min antigen-antibody reaction step and takes no more than 45 min for dozens of samples analysis was developed. The half maximal inhibitory concentration (IC 50 ) and limit of detection (LOD) of the MB-ELISA are 1.78 ng/mL and 0.13 ng/mL, respectively. And the MB-ELISA working range for corn samples analysis is from 5.0 μg/kg to 255.2 μg/kg. The recoveries for ZEN spiked corn samples ranged from 82.3 to 110.5% with coefficient of variation (CV) under 8.9%. For natural corn samples analysis, the results of MB-ELISA showed good agreement with the results of conventional direct competitive ELISA ( R 2  = 0.9742).