高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (8): 1845-1850.doi: 10.7503/cjcu20130424

• 分析化学 • 上一篇    下一篇

纳米金颗粒增强信号的电化学生物传感器用于谷胱甘肽和半胱氨酸的检测

王青, 刘卫, 羊小海, 王柯敏, 刘沛, 何磊良   

  1. 湖南大学化学生物传感与计量学国家重点实验室, 化学化工学院, 生物纳米与分子工程湖南省重点实验室, 长沙 410082
  • 收稿日期:2013-05-07 出版日期:2013-08-10 发布日期:2013-07-19
  • 通讯作者: 王柯敏,男,博士,教授,博士生导师,主要从事化学生物传感技术及纳米尺度和分子水平上获取生物化学信息的研究.E-mail:kmwang@hnu.edu.cn E-mail:kmwang@hnu.edu.cn;yangxiaohai@hnu.edu.cn
  • 基金资助:

    国家自然科学基金(批准号: 21190044, 21175035); 科技部国际合作重大项目(批准号: 2010DFB30300); 教育部"新世纪优秀人才支持计划"(批准号: NCET-09-0338)和湖南大学"青年教师成长计划"资助.

High Sensitive Glutathione and Cysteine Detection by Au Nanoparticles Enhanced Electrochemical Biosensor

WANG Qing, LIU Wei, YANG Xiao-Hai, WANG Ke-Min, LIU Pei, HE Lei-Liang   

  1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Hunan University, Changsha 410082, China
  • Received:2013-05-07 Online:2013-08-10 Published:2013-07-19
  • Contact: Kemin Wang, YANG Xiao-Hai E-mail:kmwang@hnu.edu.cn;yangxiaohai@hnu.edu.cn

摘要:

构建了一种高灵敏检测谷胱甘肽(GSH)和半胱氨酸(Cys)的新型电化学生物传感器. 先将富含T碱基的DNA1和DNA2探针分别修饰在金电极和纳米金颗粒(AuNPs)上, 再加入Hg2+, 通过形成T-Hg2+-T结构使AuNPs结合到金电极表面. 当加入GSH(或Cys)后, GSH(或Cys)可以竞争结合T-Hg2+-T结构中的Hg2+, 使AuNPs离开电极表面. 由于AuNPs上修饰的DNA探针能够静电吸附大量电活性物质六氨合钌(RuHex), 因此该过程可引起计时电量信号的显著变化, 据此实现了GSH(或Cys)的高灵敏检测. 该传感器的检出限达10 pmol/L, 比荧光法或比色法降低了2~3个数量级. 实验结果表明, 该传感器具有较好的选择性.

关键词: 电化学生物传感器, 纳米金颗粒, DNA, 谷胱甘肽, 半胱氨酸

Abstract:

Glutathione(GSH) and cysteine(Cys) are biological thiols that play vital roles in maintaining cellular functions. GSH is a thiol-group-containing tripep-tide, which has a pivotal role in maintaining the redu-cing environment in cells. Cys is a biomarker for various medical conditions, and a disease-associated physiological regulator. In this work, a high sensitive electrochemical biosensor for GSH and Cys detection was constructed using Au nanoparticles enhanced signal amplification. DNA1 and DNA2 were all poly-T DNA probes. Firstly, DNA1 was modified on the surface of electrode, and then DNA2 was modified on the AuNPs. In the presence of Hg2+, the binding of Hg2+ with thymine(T) was employed to form T-Hg2+-T structure and thus DNA2 modified AuNPs could be captured on the electrode surface. The added GSH(or Cys) could selectively coordinate with Hg2+ of the T-Hg2+-T structure, which resulted in the departure of AuNPs from the electrode surface. Since DNA modified on AuNPs could bind a large number of electric active RuHex via electrostatic interaction, the chronocoulometry signal was reduced obviously, and then GSH(or Cys) could be detected according to the change of chronocoulometry signal. The results showed that the electrochemical biosensor was highly sensitive, and it could detect as low as 10 pmol/L for GSH(or Cys). It was about 2-3 orders of magnitude lower than those of fluorescent and colorimetric methods. Moreover, eight kinds of amino acids were used as control to investigate the selectivity of this electrochemical biosensor, and the results showed that this electrochemical biosensor showed good selectivity.

Key words: Electrochemical biosensor, Au nanoparticles, DNA, Glutathione, Cysteine

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