| 雌二醇间接竞争酶联免疫吸附方法的建立 |
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| 引用本文： | 刘珒,雷红涛,孙远明,李振峰,杨金易,沈玉栋,王弘,曾道平. 雌二醇间接竞争酶联免疫吸附方法的建立[J]. 食品科学, 2012, 33(8): 146-150. DOI: 10.7506/spkx1002-6630-201208030 |
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| 作者姓名： | 刘珒 雷红涛 孙远明 李振峰 杨金易 沈玉栋 王弘 曾道平 |
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| 作者单位： | 华南农业大学食品学院，广东省食品质量安全重点实验室 |
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| 基金项目： | 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) |
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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。
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| 关 键 词： | 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 |
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| LIU Jin,LEI Hong-tao,SUN Yuan-ming,LI Zhen-feng,YANG Jin-yi,SHEN Yu-dong, WANG Hong,ZENG Dao-ping. Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Estradiol[J]. Food Science, 2012, 33(8): 146-150. DOI: 10.7506/spkx1002-6630-201208030 |
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| Authors: | LIU Jin LEI Hong-tao SUN Yuan-ming LI Zhen-feng YANG Jin-yi SHEN Yu-dong WANG Hong ZENG Dao-ping |
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| Affiliation: | (Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University,Guangzhou 510642, China) |
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| 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. |
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| Keywords: | estradiol antibody indirect competitive enzyme-linked immunosorbent assay |
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