Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

查看全文 作　　者：LI [1]WeiHua;MENG [2]Meng;HE [3]FangYang;WAN [3]YuPing;XUE [2]HuYin;LIU [1]Wei;YIN [1]WeiWei;XU [1]Jing;FENG [3]CaiWei;WANG [3]ShanLiang;LU [4]Xiao;LIU [1]JinTing;XI [2]RiMo

机构地区：[1]School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China;[2]College of Pharmacy and Tianjin key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China;[3]Beijing Wanger Biotechnology Co., Ltd., Beijing 102206, China;[4]Central Laboratory of Shandong Academy of Agricultural Sciences, Jinan 250100, China

出　　处：《Chinese Science Bulletin》索引2011年第56卷第8期,共6页

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) 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. 关键词：ELISA检测方法单克隆抗体己烯雌酚生物样品竞争间接酶联免疫吸附试验 DES

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Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

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