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