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Peroxidase immobilized on phospholipid bilayers supported on au (111) by DTT self-assembled monolayers: Application to dopamine determination

Authors

  • Maurícia B. Fritzen-Garcia,

    Corresponding author
    1. GEIMM, Departamento de Ciências Farmacêuticas, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; telephone: 55-48-3721-2212; fax: 55-48-3234-0599
    2. Ciência e Engenharia de Materiais, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil
    • GEIMM, Departamento de Ciências Farmacêuticas, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; telephone: 55-48-3721-2212; fax: 55-48-3234-0599.
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  • Vinícius C. Zoldan,

    1. LFFS, Departamento de Física, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil
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  • Inês Rosane W.Z. Oliveira,

    1. Departamento de Química, Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil
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  • Valdir Soldi,

    1. Departamento de Química, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil
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  • André A. Pasa,

    1. LFFS, Departamento de Física, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil
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  • Tânia B. Creczynski-Pasa

    Corresponding author
    1. GEIMM, Departamento de Ciências Farmacêuticas, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; telephone: 55-48-3721-2212; fax: 55-48-3234-0599
    • GEIMM, Departamento de Ciências Farmacêuticas, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; telephone: 55-48-3721-2212; fax: 55-48-3234-0599.
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Abstract

In this work, horseradish peroxidase (HRP) was immobilized on dimyristoylphosphatidylcholine (DMPC) bilayers supported on Au (111) by dithiotreitol (DTT) self-assembled monolayers and used as a nanostructured electrochemical biosensor to dopamine determination. The morphology of the phospholipid bilayers and the immobilization of HRP to these layers were characterized by atomic force microscopy (AFM). Square-wave voltammetry (SWV) experiments were done to investigate the performance of the HRP-modified electrode. The AFM images indicate that the enzyme is adsorbed at the external layer of the lipid bilayer and, although the electrical charges on the surface were not measured, the enzyme and phospholipids surface interaction occurs probably by electrostatic forces due to the pH used in the experiments. Interestingly, the present system can be used as one-shot sensor for the rapid detection of dopamine. The analytical performance of this system was linear for dopamine concentrations from 3.3 × 10−5 to 1.3 × 10−3 mol L−1 (r = 0.9997) with a detection limit of 2.0 × 10−6 mol L−1. Our results indicate that the use of HRP-DMPC bilayer system may be useful not only in developing new nanostructured materials for technological purposes, but could be very useful in fundamental studies to investigate the interactions between different micro-and macromolecules, even with soluble proteins, and lipid membranes. Biotechnol. Bioeng. 2013; 110: 374–382. © 2012 Wiley Periodicals, Inc.

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