摘要
Based on the preparation of an anti-diethylstilbestrol(DES) monoclonal antibody,a simple and convenient indirect competitive enzyme-linked immunosorbent assay(ELISA) method for DES detection has been developed.The monoclonal antibody demonstrated high sensitivity to DES with an IC50 value of 275 pg mL-1 and detection limit(LOD) of 90 pg mL-1.The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol(<7%),estrone(<0.1%),estriol(<0.1%),and diethylstilbestrol benzoate(<0.1%).Chicken,fish,shrimp,urine and bile spiked with different concentration of DES were detected by the developed method,and the recovery rates were greater than 79.5%.Intra-and inter-assay variations were about 6%.This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples.The LODs in fish/shrimp,liver,feed and urine spiked with DES were 600,600,4800 and 600 pg mL-1,respectively.These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals.
Based on the preparation of an anti-diethylstilbestrol （DES） monoclonal antibody, a simple and convenient indirect competitive enzyme-linked immunosorbent assay （ELISA） method for DES detection has been developed. The monoclonal antibody demonstrated high sensitivity to DES with an IC50 value of 275 pg mL-1 and detection limit （LOD） of 90 pg mL-1. The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol （〈7%）, estrone （〈0.1%）, estriol （〈0.1%）, and diethylstilbestrol benzoate （〈0.1%）. Chicken, fish, shrimp, urine and bile spiked with dif- ferent concentration of DES were detected by the developed method, and the recovery rates were greater than 79.5%. Intra- and inter-assay variations were about 6%. This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples. The LODs in fish/shrimp, liver, feed and urine spiked with DES were 600, 600, 4800 and 600 pg mL-1, respec- tively. These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals.
基金
supported by the Shandong Natural Science Foundation (Y2008B31)
the National High-Tech Research and Development Program of China (07AA10Z435, 2007AA06A407)
the National Natural Science Foundation of China (20675048)
the Fundamental Research Funds for the Central Universities (65011121)
