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Determination of trypsin activity using a gold electrode modified with a nanocover composed of graphene oxide and thionine

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Abstract

We report on an electrochemical method for the determination of the activity of trypsin. A multi-functional substrate peptide (HHHAKSSATGGC-HS) is designed and immobilized on a gold electrode. The three His residues in the N-terminal are able to recruit thionine-loaded graphene oxide (GO/thionine), a nanocover adopted for signal amplification. Once the peptide is cleaved under enzymatic catalysis by trypsin (cleavage site: Lys residue), the His residues leave the electrode, and the GO/thionine cannot cover the peptide-modified electrode anymore. Thus, the changes of the electrochemical signal of thionine, typically acquired at a voltage of -0.35 V, can be used to determine the activity of trypsin. A detection range of 1 × 10−4 to 1 U, with a detection limit of 3.3 × 10−5 U, can be achieved, which is better than some currently available methods. In addition, the method is highly specific, facile, and has the potential for the detection of trypsin-like proteases.

Graphene oxide was adopted as a nanocover for the development of a sensitive electrochemical method to detect the activity of trypsin.

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Acknowledgments

This work is supported by the National Natural Science Foundation of China (Grant No. 31200742).

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Authors and Affiliations

  1. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, 200444, People’s Republic of China

    Guifang Chen, Hai Shi & Fangfang Ban

  2. Department of obstetrics, The First Affiliated Hospital with Nanjing Medical University, Nanjing, 210029, People’s Republic of China

    Yuanyuan Zhang & Lizhou Sun

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  1. Guifang Chen
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  2. Hai Shi
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  3. Fangfang Ban
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  4. Yuanyuan Zhang
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  5. Lizhou Sun
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Correspondence to Lizhou Sun.

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Chen, G., Shi, H., Ban, F. et al. Determination of trypsin activity using a gold electrode modified with a nanocover composed of graphene oxide and thionine. Microchim Acta 182, 2469–2476 (2015). https://doi.org/10.1007/s00604-015-1601-x

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  • DOI: https://doi.org/10.1007/s00604-015-1601-x

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