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Nanocluster-assisted protein-film voltammetry for direct electrochemical signal acquisition

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Abstract

Acquisition of the direct electrochemical response of protein is the cornerstone for the development of the third generation of electrochemical biosensors. In this work, we developed a nanocluster-assisted protein-film voltammetry technique (NCA-PFV) which can achieve the acquisition of the electrochemical signal and maintain the activity without affecting of the protein’s structure. With this strategy, a lipid bilayer membrane is used to immobilize the membrane protein so as to maintain its natural state. Copper nanoclusters with a size smaller than most proteins are then used to function at sub-protein scale and to mediate the electron hopping from the electroactive center of the electrode. As a model, the direct electrochemical signal of cyclooxygenase (COX) is successfully obtained, with a pair of well-defined redox peaks located at −0.39 mV and −0.31 mV, which characterize the heme center of the enzyme. Its catalytic activity towards the substrate arachidonic acid (AA) is also retained. The detection range for AA is 10–1000 μM and the detection limit is 2.4 μM. Electrochemical monitoring of the regulation of the catalytic activity by an inhibitor DuP-697 is also achieved. This work provides a powerful tool for the fabrication of enzyme-based electrochemical biosensors, and is also of great significance for promoting the development and application of next-generation electrochemical biosensors.

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Authors and Affiliations

  1. Clinical Laboratory, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, 570208, China

    Hui Huang

  2. State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, 570228, China

    Wenxing Li

  3. College of Resources and Environmental Engineering, Shandong University of Technology, Zibo, 255049, China

    Xiaomei Yu

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  1. Wenxing Li
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  2. Xiaomei Yu
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  3. Hui Huang
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Correspondence to Hui Huang.

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This work was supported by the Scientific Research Projects of Health Commission in Hainan Province [1901031041A2001], and the authors declare no competing financial or non-financial interests. The human plasma study was conducted according to the principles expressed in the Declaration of Helsinki and was approved by the Ethical Committee of Haikou People’s Hospital. All subjects provided written informed consent to participate.

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Li, W., Yu, X. & Huang, H. Nanocluster-assisted protein-film voltammetry for direct electrochemical signal acquisition. Anal Bioanal Chem 413, 1665–1673 (2021). https://doi.org/10.1007/s00216-020-03130-5

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  • DOI: https://doi.org/10.1007/s00216-020-03130-5

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