雌二醇单克隆抗体制备及间接竞争ELISA检测方法的建立

发布时间：2018-02-27 17:24

本文关键词： 雌二醇单克隆抗体间接竞争ELISA　出处：《中国畜牧兽医》2017年10期 　论文类型：期刊论文

【摘要】：为建立敏感、特异、快速的雌二醇(estradiol,E2)残留免疫检测方法,本研究利用雌二醇人工抗原(E2-BSA)免疫BALB/c小鼠,应用淋巴细胞杂交瘤技术制备特异性雌二醇单克隆抗体,利用高效价、高特异性单克隆抗体建立间接竞争ELISA(ic ELISA)方法。结果显示,试验成功筛选获得一株稳定分泌抗雌二醇抗体的杂交瘤细胞株(3H3),抗体效价可达1∶320 000,利用3H3腹水抗体优化间接竞争ic ELISA反应条件,检测抗体的敏感度,IC50为1.636 ng/m L,IC20~IC80线性范围为0.202~13.281 ng/m L。雌二醇腹水抗体与雌三醇、乙炔雌二醇的交叉反应率分别达0.31%和0.25%,而与雌酮、戊酸雌二醇、苯甲酸雌二醇、炔雌醚、己烯雌酚、壬基酚的交叉反应率均0.1%。结果表明,利用本研究制备的单克隆抗体建立雌二醇间接竞争ELISA检测方法,可满足食品中雌二醇残留高灵敏度的检测要求。
[Abstract]:In order to establish a sensitive, specific and rapid method for the detection of estradiolium estradiolium E2), BALB/c mice were immunized with estradiol artificial antigen E2-BSA. specific estradiol monoclonal antibodies were prepared by lymphocyte hybridoma technique. An indirect competitive ELISA(ic Elisa method was established with highly specific monoclonal antibodies. The results showed that, A hybridoma cell strain with stable secretion of anti-estradiol antibody was successfully screened. The titer of the antibody was 1: 320 000. The indirect competitive ic ELISA reaction conditions were optimized by using 3H3 ascites antibody. The sensitivity of IC50 was 1.636 ng/m / L, and the linear range of IC20 / IC80 was 0.31% ng/m / L, 13.281 ng/m / L, respectively. The cross reaction rates of estradiol ascites antibody with estriol and ethylestradiol were 0.31% and 0.25, respectively. The cross reaction rates of nonylphenol were 0.1%. The results showed that the indirect competitive ELISA assay of estradiol with monoclonal antibody prepared in this study could meet the requirements of high sensitivity for the detection of estradiol residues in food.
【作者单位】：温州市农业科学研究院动物科学研究所;温州科技职业学院动物科学学院;
【基金】：浙江省公益性技术研究农业项目(2014C32046) 温州市公益性社会发展科技项目(S20150026)
【分类号】：S859.84

【相似文献】