Development of pyruvate oxidase-based amperometric biosensor for pyruvate determination

Authors

  • D. V. Knyzhnykova Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author
  • Ya. V. Topolnikova Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author
  • I. S. Kucherenko Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author
  • O. O. Soldatkin Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author

DOI:

https://doi.org/10.7124/bc.00096C

Keywords:

pyruvate, pyruvate oxidase, amperometric biosensor

Abstract

Aim. The development and optimization of the amperometric biosensor for pyruvate determina-tion. Methods. Immobilized pyruvate oxidase was used as a biorecognition element of the biosen-sor, a platinum disc electrode- as an electrochemical transducer. Results. Different variants of immobilization of pyruvate oxidase were tested and the optimal one was chosen for the creation of a biorecognition element of the biosensor. Optimal concentrations of cofactors for the best per-formance of the pyruvate oxidase-based biosensor were selected. The developed biosensor dem-onstrated a high sensitivity to pyruvate and wide linear range of work. High selectivity of the pro-posed biosensor towards electrically active substances and other substrates present in real samples was shown. The biosensor is characterized by high signal reproducibility and operational stability over two weeks. Conclusions. The highly selective amperometric biosensor for determination of pyruvate in biological samples has been developed. Its analytical characteristics are studied. The biosensor can be further used for the pyruvate analysis in blood serum.

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Published

2018-02-28

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Section

Molecular and Cell Biotechnologies