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Anal Chem. 2010 May 1;82(9):3588-96. doi: 10.1021/ac100621r.

Electrochemical approach for detection of extracellular oxygen released from erythrocytes based on graphene film integrated with laccase and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid).

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  • 1Jiangsu Key Laboratory of Biofunctional Materials, Laboratory of Electrochemistry, College of Chemistry and Environmental Science, Nanjing Normal University, Nanjing 210097, PR China.


This work develops a novel electrochemical approach for detection of the extracellular oxygen released from human erythrocytes. The sensing is based on the bioelectrocatalytic system of graphene integrated with laccase (Lac) and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) toward the reduction of oxygen. ABTS and laccase are assembled on the surface of graphene, which is synthesized by a chemistry route, utilizing the pi-pi and electrostatic interactions of these components. Transmission electron microscopy (TEM), atomic force microscopy (AFM), and FT-IR spectroscopy demonstrate that graphene has been successfully synthesized, and ABTS and laccase have been effectively assembled on a graphene surface with the formation of Lac-ABTS-graphene hybrid. The voltammetric results indicate that ABTS can be used as a redox mediator when it is in immobilized form. The hybrid deposited on the glassy carbon (GC) electrode is demonstrated to be a good bioelectrocatalyst for the reduction of oxygen with inherent enzyme activity, accepted stability, high half-wave potential (ca.670 mV vs NHE), and unimpeded electrical communication to the copper redox sites of laccase. Therefore, this study has not only established a novel approach of detection of extracellular oxygen but also provided a general route for fabricating a graphene-based biosensing platform via assembling enzymes/proteins on a graphene surface.

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