Investigation of diethylstilbestrol residue level in human urine samples by a specific monoclonal antibody
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Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people’s health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC50 of this method was 3.33 ng/mL. The LOD and LOQ were 0.16 and 0.54 ng/mL. Linear range of the standard curve was from LOD to 12.50 ng/mL. There was no cross-reactivity with two kinds of estrogens and two structural analogs with DES. Five hundred seventy-six urine samples were analyzed by the indirect competitive ELISA method, and the detection rate was 98.78%. The mean concentration and geometric mean were 4.70 and 3.50 ng/mL. The indirect competitive ELISA method based on monoclonal antibody was sensitive and reliable for the detection of DES in human urine samples. The results warned us to pay more attention to human health and food safety.
KeywordsDiethylstilbestrol Monoclonal antibody ELISA Human urine samples Residue level Human health
This work was supported by the State Key Program of National Natural Science of China (Grant No. 81430081), the grant (No. 2012ZX09506001-004) from the National Major Projects for Science and Technology Development of Ministry Science and Technology of China, and the Fundamental Research Funds for the Central Universities.
Compliance with ethical standards
All experiments on mice were performed according to the ethics committee of the Experimental Animal Center of Zhejiang University.
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