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

Abstract 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.