雌二醇间接竞争酶联免疫吸附方法的建立.PDF

146 2012, Vol. 33, No. 08 食品科学 ※分析检测 刘 珒，雷红涛 * ，孙远明，李振峰，杨金易，沈玉栋，王 弘，曾道平 (华南农业大学食品学院，广东省食品质量安全重点实验室，广东 广州 510642) 建立食品中雌二醇免疫分析方法。将雌二醇进行结构改造合成半抗原并经过核磁氢谱和红外光谱验证，再 采用活泼酯法与载体 BSA 和 OVA 偶联制备人工抗原，免疫兔子制备多克隆抗体，经过反应条件优化建立间接竞争 酶联免疫吸附检测方法。结果表明：IC50 为 3.9ng/mL ，检测限为0.3ng/mL ，交叉反应率均小于 1.21%。检测方法 具有高灵敏度和特异性，可应用于食品中雌二醇检测试剂盒的研制。 雌二醇；抗体；间接竞争酶联免疫检测 Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol LIU Jin ，LEI Hong-tao* ，SUN Yuan-ming，LI Zhen-feng ，YANG Jin-yi ，SHEN Yu-dong， WANG Hong ，ZENG Dao-ping (Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China) Abstract ：In order to establish a simple and fast assay for the detection of estradiol in foods, estradiol hapten was synthesized and identified by nuclear magnetic resonance (NMR) and infrared spectroscopy. The hapten was activated by DCC/NHS ester method and covalently coupled to carrier proteins (bovine serum albumin and ovalbumin). Polyclonal antibodies were obtained from immunized rabbits. The optimal reaction conditions of indirect competitive enzyme-linked immunosorbent assay (icELISA) were established. The IC50 and LOD of the developed method were 3.9 ng/mL and 0.3 ng/mL, respectively. The cross-reactivity with one of 8 structural analogues of estradiol was less than 1.21%. High sensitivity and specificity will contribute to the development of commercial ELISA kits for the detection of estradiol in foods. Key words ：estradiol ；antibody ；indirect competitive enzyme-linked immunosorbent assay 中图分类号：TS201.3 文献标识码：A 文章编号：1002-6630(2012)08-0146-05 为了提高瘦肉生长率和饲料利用率