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

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Abstract

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.

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.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 31200742, 81272601, 30973477), the “Chen Guang” project of Shanghai Municipal Education Commission and Shanghai Education Development Foundation (No. 10CG42), the Innovation Program of Shanghai Municipal Education Commission (No. 12YZ004) and the Leading Academic Discipline Project of Shanghai Municipal Education Commission (J50108).

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Authors and Affiliations

  1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210093, Peoples Republic of China

    Zonghuang Ye & Genxi Li

  2. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, 200444, Peoples Republic of China

    Zonghuang Ye, Bin Zhang, Xiaoli Zhu & Genxi Li

  3. Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, Peoples Republic of China

    Yucai Yang & Zhaoxia Wang

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  1. Zonghuang Ye
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  2. Bin Zhang
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  3. Yucai Yang
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  4. Zhaoxia Wang
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  5. Xiaoli Zhu
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  6. Genxi Li
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Correspondence to Zhaoxia Wang or Xiaoli Zhu.

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Ye, Z., Zhang, B., Yang, Y. et al. Electrochemical biosensor for the nuclear factor kappa B using a gold nanoparticle-assisted dual signal amplification method. Microchim Acta 181, 139–145 (2014). https://doi.org/10.1007/s00604-013-1080-x

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  • DOI: https://doi.org/10.1007/s00604-013-1080-x

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