氨基纳米磁球免疫电化学法检测甲胎蛋白的研究  

Immuno-electrochemical Detection of Alpha-fetoprotein Based on Nano-magnetic Sphere with Amino Group

作　　者：程琼[1] 彭图治[1] 刘爱丽[1] 何亮 

机构地区：[1]浙江大学西溪校区化学系 [2]杭州埃夫朗生物技术有限公司,杭州310014

出　　处：《分析化学》2005年第8期1068-1072,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.29975024;20275034);浙江省分析测试基金(No.04062);浙江省重点科技计划项目(No.2003C21024)资助项目

摘　　要：提出了一种利用纳米磁球免疫分离,进行酶催化电化学检测甲胎蛋白的新方法.在自制含有活性氨基的纳米磁球表面,用戊二醛固化甲胎蛋白(AFP)抗体,利用免疫夹心反应原理,捕获溶液中的AFP抗原和标记有辣根过氧化物酶的AFP抗体.在外加磁场的作用下,抗体-抗原结合物从样品溶液中分离,在含有邻氨基苯酚和过氧化氢的底液中,生成具有电活性的化合物3-氨基吩呃嗪,用灵敏的示差脉冲伏安法测定.响应电流与AFP抗原浓度分别在1～5和 5～400 μg/L范围内呈线性关系, 检出限达0.5 μg/L.实验表明,该方法具有分离效率高、测定时间短、抗干扰性强等优点,尤其适用于分析复杂生物样品.应用此法于人血清样品中AFP的检测,灵敏度显著高于酶联免疫吸附法.A novel electrochemical method of enzyme catalysis using magnetic separation with nano-magnetic spheres was developed for rapid and sensitive detection of alpha-fetoprotein （AFP）. The surface of the amino nano-magnetic sphere was covalently bound with AFP antibody by glutaraldehyde, and then AFP antigen and AFP antibody labeled with horseradish peroxide enzyme were captured on the surface respectively. The sphere with “sandwich” complex was separated from sample solution by a magnetic field and the enzyme labeled at the complex catalyzed the reaction of 2-amino hydroxybenzene and hydrogen peroxide in the buffer solution.The electroactive product of 3-amino phenoxazine was detected using differential pulse voltammetry. The peak current was linear with the concentration of AFP antigen in the range of 1 - 5 and 5 - 400μg/L respectively under the optimum conditions. The detection limit was found to be 0.5μg/L. The results showed the method possessedthe advantages of high separation efficency, short measureing time and good selectivity, which were well suited for assay of complex bio-compounds. The method has been used to detect the concentration of AFP in human serum samples and the sensitivity was much higher than that of enzyme-linked immunosorbnent assay（ELISA） used in diagnostic analysis.

关 键 词：氨基纳米磁球 免疫电化学 检测 甲胎蛋白 抗体 

分 类 号：R319[医药卫生—基础医学]