Development of a broad-spectrum monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for screening of androgens in animal edible tissues

•A sensitive and broad-spectrum mAb against 4 androgens was obtained.•Molecular modeling was used to hapten design and antibody analyze.•A mAb-based ic-ELISA was developed for 4 androgens in 11 various samples. Androgens efficiently promote skeletal muscle growth and protein synthesis but causes neurotoxicity, genotoxicity and carcinogenicity, which had been prohibited applying in livestock. In this study, a broad-spectrum monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed for rapidly screening of androgens in various animal tissues. Primarily, a group-specific mAb, 4D12, was produced and showed IC50 values for nortestosterone, methyltestosterone, testosterone and trenbolone of 2.2, 0.3, 1.6 and 4.4 μg L−1, respectively. The limits of detection and limits of quantification of this method for 4 androgens in 11 kinds of animal edible tissues ranged from 37.2 to 697.8 ng L−1 and 70.0 to 1524.0 ng L−1, respectively. The recoveries of the 11 kinds of samples spiked with 4 androgens were in the range from 65.0% to 106.6%, with coefficients of variation below 16.9%. A good correlation (R2 = 0.998) was shown between the ic-ELISA and HPLC-MS/MS results for beef.