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| 雌二醇单克隆抗体制备及间接竞争ELISA检测方法的建立 | |
| 引用本文： | 白宇,胡景炎,许永鹏,何杰,金俊杰,李培德,刘素贞.雌二醇单克隆抗体制备及间接竞争ELISA检测方法的建立[J].中国畜牧兽医,2017,44(10):3100-3105. |
| 作者姓名： | 白宇 胡景炎 许永鹏 何杰 金俊杰 李培德 刘素贞 |
| 作者单位： | 1. 温州市农业科学研究院动物科学研究所, 温州 325006; 2. 温州科技职业学院动物科学学院, 温州 325006 |
| 基金项目： | 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） |
| 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（icELISA）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icELISA反应条件，检测抗体的敏感度，IC50为1.636 ng/mL，IC20~IC80线性范围为0.202~13.281 ng/mL。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。 |
| 关 键 词： | 雌二醇 单克隆抗体 间接竞争ELISA |
| 收稿时间： | 2017-04-12 |
Preparation of Monoclonal Antibody and Establishment of Indirect Competitive ELISA of Estradiol |
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| BAI Yu,HU Jing-yan,XU Yong-peng,HE Jie,JIN Jun-jie,LI Pei-de,LIU Su-zhen.Preparation of Monoclonal Antibody and Establishment of Indirect Competitive ELISA of Estradiol[J].China Animal Husbandry & Veterinary Medicine,2017,44(10):3100-3105. | |
| Authors: | BAI Yu HU Jing-yan XU Yong-peng HE Jie JIN Jun-jie LI Pei-de LIU Su-zhen |
| Institution: | 1. Institute of Animal Science, Wenzhou Academy of Agricultural Sciences, Wenzhou 325006, China; 2. College of Animal Science, Wenzhou Vocational College of Science and Technology, Wenzhou 325006, China |
| Abstract: | In order to establish a sensitive,specific and rapid method for the detection of estradiol (E2) residues, an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibody (MAb) for E2 was developed. BALB/c mice were immunized by E2-BSA and cell fusion technology was employed to screen hybridoma cell lines. One hybridoma cell line (3H3) was isolated, which produced monoclonal antibody that could binding E2. Under the optimized conditions, the icELISA based on 3H3 for E2 showed a half maximum inhibition concentration (IC50) values of 1.636 ng/mL and detection ranges of 0.202 to 13.281 ng/mL with cross-reactivities for estriol and ethinyloestradiol of 0.31% and 0.25%, respectively, and negligible cross-reactivities with other E2 analogs including estrone, estradiol valerate, estradiol benzoate, quinestrol, diethylstilbestrol and nonylphenol. The results demonstrated that the developed method could meet the requirements of high sensitivity detection of E2 residue in food samples. |
| Keywords: | estradiol monoclonal antibody indirect competitive ELISA |
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