己烯雌酚特异性抗体的制备及鉴定  

Preparation and Identification of the Specific Antibody against Diethylstilbestrol

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作  者:邓省亮[1,2] 李平[1] 刘洪斌[1] 程玛丽[3] 于洪侠[1] 陈刚[1] 杨曙明[1] 

机构地区:[1]中国农业科学院农业质量标准与检测技术研究所,北京100081 [2]江西省科学院微生物研究所,南昌330029 [3]北京农学院动物科学技术学院,北京102206

出  处:《中国生物工程杂志》2011年第6期106-110,共5页China Biotechnology

基  金:国家"863"计划(2007AA10Z438);科技部科技支撑专项(2009BADB7B07)资助项目

摘  要:利用4-溴丁酸乙酯对小分子半抗原己烯雌酚(DES)进行活化,引入羧基活性基团,应用活泼酯法将其与牛血清白蛋白(BSA)偶联,合成DES-CP-BSA完全抗原,免疫新西兰长耳白兔,制备特异性抗体。结果显示:成功制备了DES完全抗原,且由此获得了特异性的DES抗体,效价达1.28×105,与己烷雌酚、双烯雌酚的交叉反应分别为9.3%和1.1%,与雌二醇无交叉反应。在此基础上建立了间接竞争ELISA检测方法,IC50为7.0 ng/ml,检测限达0.3 ng/ml,可以满足动物性食品中DES残留检测的需要。Diethylstilbestrol was activated by ethyl 4-bromobutyrate to introduce carboxyl group and then conjugated to BSA via EDC/NHS as immunogen, New Zealand rabbits were immunized and antisera were prepared. The results showed that complete antigen was successfully synthesized and specific antibody was generated, with titer of 1.28 x 105 the antisera cross-reactivity with hexestrol (9.34%), dienestrol ( 1.1% ) and no cross-reactivity with 17[3-estradiol. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed for determination of DES. In the linear range between 1 and 30 ng/ml, IC50 of 7 ng/ml and a detection limit of 0. 3 ng/ml for DES were reached. Therefore the assay as screening method can meet the demand of DES residue detection for animal food products.

关 键 词:己烯雌酚 特异性抗体 ELISA 

分 类 号:Q819[生物学—生物工程]

 

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