雌二醇间接竞争酶联免疫吸附方法的建立  

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol

作　　者：刘珒[1] 雷红涛[1] 孙远明[1] 李振峰[1] 杨金易[1] 沈玉栋[1] 王弘[1] 曾道平[1] 

机构地区：[1]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

出　　处：《食品科学》2012年第8期146-150,共5页Food Science

基　　金：国家自然科学基金面上项目(30700663);教育部高等学校博士点基金项目(20114404130002);广州市科技计划项目(11C12100483);国家公益性行业(农业)科研专项(201003008-08)

摘　　要：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。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.

关 键 词：雌二醇 抗体 间接竞争酶联免疫检测 

分 类 号：TS201.3[轻工技术与工程—食品科学]