Electrochemical biosensor for the nuclear factor kappa B using a gold nanoparticle-assisted dual signal amplification method

详细信息    查看全文

• 作者：Zonghuang Ye (1) (2)
  Bin Zhang (2)
  Yucai Yang (3)
  Zhaoxia Wang (3)
  Xiaoli Zhu (2)
  Genxi Li (1) (2)
  
• 关键词：NF ; κB ; Transcription factor ; Nicking endonuclease ; Gold nanoparticles ; Electroanalysis
• 刊名：Microchimica Acta
• 出版年：2014
• 出版时间：January 2014
• 年：2014
• 卷：181
• 期：1-2
• 页码：139-145
• 全文大小：262 KB
• 作者单位：Zonghuang Ye (1) (2)
  Bin Zhang (2)
  Yucai Yang (3)
  Zhaoxia Wang (3)
  Xiaoli Zhu (2)
  Genxi Li (1) (2)
  
  1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210093, Peoples Republic of China
  2. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, 200444, Peoples Republic of China
  3. Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, Peoples Republic of China
  
• ISSN：1436-5073

文摘

Nuclear factor kappa B (NF-κB) is a transcription factor that plays a central role in the signaling pathway and network of gene regulation. The dysregulation of NF-κB signaling has been implicated in the pathogenesis of a number of diseases. We report on a dual amplification strategy for the highly sensitive electrochemical sensing of NF-κB by means of a nicking endonuclease-assisted amplification reaction (NEAR). The quantity of the DNA obtained is subsequently determined by applying a gold nanoparticle-assisted electrochemical amplification step. This represents the first example of a combination of NEAR and a dual amplification strategy for the detection of a transcription factor. Experimental results show that the electrochemical signal generated by the redox probe (the ruthenium(III) hexammine complex) can be related to the concentration of NF-κB. The response of the electrode is linearly related to the concentration of NF-κB in the 100?pM to 10?nM range, with a detection limit as low as 80?pM. Figure An electrochemical biosensor for NF-κB with dual signal amplification is achieved. It was based on nicking endonuclease-assisted amplification reaction (NEAR) and gold nanoparticle-assisted electrochemical amplification (AuNP-EA). The method can be extended for the detection of universal DNA-binding proteins.