化学学报 ›› 2011, Vol. 69 ›› Issue (10): 1211-1216. 上一篇    下一篇

研究论文

金增强多重胶体金标记树枝状复合物放大的电化学免疫分析新方法

王潇蕤*, 李继睿   

  1. (湖南化工职业技术学院应用化学系 株洲 412004)
  • 投稿日期:2010-09-16 修回日期:2010-11-25 发布日期:2011-01-10
  • 通讯作者: 王潇蕤 E-mail:xr-wang@hotmail.com

Electrochemical Immunoassay Based on Gold-Enhanced Dendritic Complex Loaded with Multiple Colloidal Gold Labels

WANG Xiao-Rui, LI Ji-Rui   

  1. (Department of Applied Chemistry, Hunan Chemical Industry Vocation Technology Institute, Zhuzhou 412004)
  • Received:2010-09-16 Revised:2010-11-25 Published:2011-01-10

报道了一种电化学免疫分析新方法用于痕量(amol/L级)蛋白质的高灵敏检测. 该法采用了两级信号放大策略. 先以金标蛋白A与金标抗体形成的树枝状复合物作为一级信号放大, 随后使用金增强溶液催化捕获在此复合物上的大量的胶体金纳米颗粒的增大以实现二级信号放大. 这种两级信号放大步骤进而结合灵敏的金属溶出伏安分析法以有效增强抗原-抗体免疫反应响应. 实验还通过引入免疫复合物的磁分离操作来降低非特异性吸附的影响. 结果表明, 在优化的实验条件下, 开发的新方法在用于人IgG模型分析物测定时的检测下限可低至5.2 fg•mL-1或34.7 amol/L (S/N=3). 这种基于胶体金的金属免疫分析新技术有望用于临床诊断、环境监测、生化防预等诸多领域中目标蛋白质的高灵敏检测.

关键词: 电化学免疫分析, 信号放大, 多重胶体金标记, 溶出伏安分析

A new electrochemical immunoassay protocol for attomolar detection of proteins has been developed by the use of the dendritic immunocomplex of protein A- and antibody-modified colloidal gold labels for the primary amplification and the catalytic enlargement of these multiple colloidal gold nanoparticles for the secondary amplification. Such use of double amplification procedure is combined with a sensitive stripping analysis of the dissolved gold tags to achieve a dramatic enhancement of the antigen-antibody binding response. The efficient magnetic separation of the immunocomplex can additionally minimize nonspecific adsorption effects. Factors affecting the performance have been optimized. Such amplified electrical transduction is demonstrated for a near-homogeneous suspension immunoassay of the human IgG model analyte with the detection limit of 5.2 fg•mL-1 or 34.7 amol/L (S/N=3). This new colloidal gold-based metalloimmunoassay approach holds great promise for highly sensitive detection of target proteins in various fields such as biodiagnostics, environmental monitoring and biodefense.

Key words: electrochemical immunoassay, signal amplification, multiple colloidal gold label, stripping analysis