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Fast and Easy Enzyme Immobilization by Photoinitiated Polymerization for Efficient Bioelectrochemical Devices
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    Fast and Easy Enzyme Immobilization by Photoinitiated Polymerization for Efficient Bioelectrochemical Devices
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    Université de Bordeaux, Centre de Recherche Paul Pascal, CNRS UPR 8641, 115 avenue Schweitzer, 33600 Pessac, France
    Université de Bordeaux, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France
    CNRS, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France
    § Groupe Nanosystèmes Analytiques, Institut des Sciences Moléculaires, CNRS UMR 5255, Université Bordeaux 1, ENSCPB, 16 avenue Pey-Berland, 33607 Pessac, France
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    Analytical Chemistry

    Cite this: Anal. Chem. 2011, 83, 7, 2824–2828
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    https://doi.org/10.1021/ac200297r
    Published March 15, 2011
    Copyright © 2011 American Chemical Society

    Abstract

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    Immobilization and electrical wiring of enzymes is of particular importance for the elaboration of efficient biosensors and can be cumbersome. Here, we report a fast and easy protocol for enzyme immobilization, and as a proof of concept, we applied it to the immobilization of bilirubin oxidase, a labile enzyme. In the first step, bilirubin oxidase is mixed with a redox hydrogel “wiring” the enzyme reaction centers to electrodes. Then, this adduct is covered by an outer layer of PEGDA made by photoinitiated polymerization of poly(ethylene-glycol) diacrylate (PEGDA) and a photoclivable precursor, DAROCUR. This two-step protocol is 18 times faster than the current state-of-the-art protocol and leads to currents 25% higher. In addition, the outer layer of PEGDA acts as a protective layer increasing the lifetime of the electrode by 100% when operating continuously for 2000 s and by 60% when kept in dry state for 24 h. This new protocol is particularly appropriate for labile enzymes that quickly denaturate. In addition, by tuning the ratio PEGDA/DAROCUR, it is possible to make the enzyme electrodes even more active or more stable.

    Copyright © 2011 American Chemical Society

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    Supporting Information

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    Details on the influence of the illumination and of the PEGDA/DAROCUR on the BOD activity and optimization of the PEGDA/DARCOCUR ratio, dilution factor, and illumination time. This material is available free of charge via the Internet at http://pubs.acs.org.

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    Cited By

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    This article is cited by 11 publications.

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    2. Nicolas Mano, Anne de Poulpiquet. O2 Reduction in Enzymatic Biofuel Cells. Chemical Reviews 2018, 118 (5) , 2392-2468. https://doi.org/10.1021/acs.chemrev.7b00220
    3. Ping Yu, Heng Zhou, Hanjun Cheng, Qin Qian, and Lanqun Mao . Rational Design and One-Step Formation of Multifunctional Gel Transducer for Simple Fabrication of Integrated Electrochemical Biosensors. Analytical Chemistry 2011, 83 (14) , 5715-5720. https://doi.org/10.1021/ac200942a
    4. Matthew S. Brown, Matthew Kelly, Ahyeon Koh. Electrochemical analysis of biological fluids. 2024, 437-483. https://doi.org/10.1016/B978-0-323-99147-6.00003-X
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    7. Serge Cosnier, Michael Holzinger, Alan Le Goff. Recent Advances in Carbon Nanotube-Based Enzymatic Fuel Cells. Frontiers in Bioengineering and Biotechnology 2014, 2 https://doi.org/10.3389/fbioe.2014.00045
    8. Marine Cadet, Xavier Brilland, Sébastien Gounel, Frédéric Louerat, Nicolas Mano. Design of a Highly Efficient O 2 Cathode Based on Bilirubin Oxidase from Magnaporthe oryzae. ChemPhysChem 2013, 14 (10) , 2097-2100. https://doi.org/10.1002/cphc.201300027
    9. Emmanuel Suraniti, Solange Vivès, Seiya Tsujimura, Nicolas Mano. Designing Thin Films of Redox Hydrogel for Highly Efficient Enzymatic Anodes. Journal of The Electrochemical Society 2013, 160 (6) , G79-G82. https://doi.org/10.1149/2.072306jes
    10. Emmanuel Suraniti, Margot Abintou, Fabien Durand, Nicolas Mano. Heat and drying time modulate the O2 reduction current of modified glassy carbon electrodes with bilirubin oxidases. Bioelectrochemistry 2012, 88 , 65-69. https://doi.org/10.1016/j.bioelechem.2012.06.002
    11. Milena Milutinovic, Emmanuel Suraniti, Vincent Studer, Nicolas Mano, Dragan Manojlovic, Neso Sojic. Photopatterning of ultrathin electrochemiluminescent redox hydrogel films. Chemical Communications 2011, 47 (32) , 9125. https://doi.org/10.1039/c1cc12724a

    Analytical Chemistry

    Cite this: Anal. Chem. 2011, 83, 7, 2824–2828
    Click to copy citationCitation copied!
    https://doi.org/10.1021/ac200297r
    Published March 15, 2011
    Copyright © 2011 American Chemical Society

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