Development of a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for screening of diethylstilbestrol in animal-derived foods

开发一种基于单克隆抗体的间接竞争酶联免疫吸附试验,用于筛查动物源性食品中的己烯雌酚

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Abstract

Diethylstilbestrol (DES), a synthetic non-steroid estrogen, it has been prohibited from being added to animal feed for any purposes. Herein, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a specific monoclonal antibody (mAb) was developed for the rapid screening of DES. Primarily, conjugates of mono-O-3-carboxypropyl diethylstilbestrol with keyhole limpet hemocyanin were used to raise a specific mAb, 1B7, which had IC(50) value for DES of 213.0 ng L(-1). The limits of detection and limits of quantification value for DES in animal-derived foods ranged from 68.1 to 103.1 ng L(-1) and 100.8-192.7 ng L(-1), respectively. The DES recovery ranged from 70.1 % to 103.1 %, with coefficients of variation below 13.9 %. A positive correlation (R(2) = 0.997) was observed between the results of ic-ELISA and HPLC-MS/MS for milk. In order to inspect its detection effect, milk and animal-derived foods were chosen as the testing object. The results showed that this ic-ELISA method (specific mAb, 1B7) can effectively examine for DES residues.

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