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雌二醇间接竞争酶联免疫吸附方法的建立 被引量:7

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol
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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.
出处 《食品科学》 EI CAS CSCD 北大核心 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)
关键词 雌二醇 抗体 间接竞争酶联免疫检测 estradiol antibody indirect competitive enzyme-linked immunosorbent assay
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参考文献16

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