高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (5): 886.

• 研究论文 • 上一篇    下一篇

基于铜增强纳米金标记物结合磁分离的溶出伏安免疫分析法用于蛋白质的高灵敏测定

沈广宇, 冯辉, 郝爱平   

  1. 湖南文理学院化学化工学院, 常德 415000
  • 收稿日期:2008-12-23 出版日期:2009-05-10 发布日期:2009-05-10
  • 通讯作者: 沈广宇, 女, 博士, 讲师, 主要从事先进功能纳米材料的制备及其生物传感应用研究, E-mail: sgyrab@yahoo.com.cn

Anodic Stripping Voltammetric Immunoassay Based on Coupling Copper-enhanced Nanogold Tags with Magnetic Separation for Highly Sensitive Detection of Proteins

SHEN Guang-Yu*, FENG Hui, HAO Ai-Ping   

  1. College of Chemistry and Chemical Engineering, Hunan University of Arts and Science, Changde 415000, China
  • Received:2008-12-23 Online:2009-05-10 Published:2009-05-10
  • Contact: SHEN Guang-Yu, E-mail: sgyrab@yahoo.com.cn

摘要:

应用铜原位化学放大纳米金颗粒的信号增强特性, 并结合磁分离技术, 提出了一种高灵敏的溶出伏安免疫分析方法. 实验中以人IgG为模式蛋白质, 将抗体修饰的SiO2@Fe3O4核壳型磁性纳米颗粒和纳米金标抗体悬浊液混合, 用以均相免疫识别人IgG, 借助外加磁场分离纯化, 在免疫复合物悬浊液中加入铜增强试剂进行沉积放大反应, 再将铜用稀硝酸溶解并进行溶出伏安分析检测. 结果表明, 与基于固相反应的金属免疫分析法相比, 所提出的基于均相反应和磁分离原理的方法具有操作简单、分析时间短等优点. 该方法显示出明显增强的人IgG检测性能, 其线性检测范围为01~1000 ng/mL, 检出限为73 pg/mL. 此外, 将其用于实际样品的回收率测定, 结果令人满意.

关键词: 纳米金, 铜增强信号放大, 磁分离, 金属免疫分析

Abstract:

A novel and highly sensitive anodic stripping voltammetric immunoassay was proposed on the basis of chemical copper-enhanced gold nanoparticle(nanogold) tags in vitro as the amplification approach with the aid of magnetic separation technique. In a homogeneous bulk solution phase, the human IgG(hIgG) as a model of test system was captured by goat anti-human IgG antibody(anti-hIgG)-conjugated SiO2@Fe3O4 magnetic nanoparticle and anti-hIgG-conjugated nanogold, followed by the copper metal dissolution in diluted nitric acid solution for anodic stripping voltammetry measurement. Comparing with the traditional solid interface reaction-based metal immunoassay, this novel one which introduce homogeneous reaction strategy combined with magnetic separation can provide some advantages, including easy of use, shorter analytical time, and especially amplified capability of detecting hIgG. It can quantitatively determine hIgG in the concentration range of 0.1—1000 ng/mL, with a detection limit about 73 pg/mL.

Key words: Nanogold, Copper-enhanced amplification, Magnetic separation, Metal immunoassay

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