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A novel third generation uric acid biosensor using uricase electro-activated with ferrocene on a Nafion coated glassy carbon electrode
University of Calcutta, India.
University of Calcutta, India.
Linköping University, Department of Physics, Chemistry and Biology, Biosensors and Bioelectronics. Linköping University, The Institute of Technology.ORCID iD: 0000-0002-1815-9699
2015 (English)In: Bioelectrochemistry, ISSN 1567-5394, E-ISSN 1878-562X, Vol. 102Article in journal (Refereed) Published
Abstract [en]

A new uric acid biosensor was constructed using ferrocene (Fc) induced electro-activated uricase (UOx) deposited within Nation (Naf) on glassy carbon electrode (GCE). Electro-activation of UOx was successfully achieved by cyclic voltammetry through the electrostatic interaction of Fc with Trp residues within the hydrophobic pockets in UOx. The Naf/UOx/Fc composite was characterised by AFM, FTIR and EDX to ensure proper immobilisation. The interaction of Fc with the enzyme was analysed by Trp fluorescence spectroscopy and the alpha-helicity of the protein was measured by CD spectropolarimetry. The charge transfer resistance (R-ct), calculated from electrochemical impedance spectroscopy, for the modified sensor was lowered (1.39 k Omega) and the enzyme efficiency was enhanced by more than two fold as a result of Fc incorporation. Cyclic voltammetty, differential pulse voltammetty and amperometry all demonstrated the excellent response of the Naf/UOx/Fc/GCE biosensor to uric acid. The sensor system generated a linear response over a range of 500 nM to 600 mu M UA, with a sensitivity and limit of detection of 1.78 mu A mu M-1 and 230 nM, respectively. The heterogeneous rate constant (k(s)) for UA oxidation was much higher for Naf/UOx/Fc/GCE (1.0 x 10(-4) cm s(-1)) than for Naf/UOx/GCE (8.2 x 10(-5) cm s(-1)). Real samples, i.e. human blood, were tested for serum UA and the sensor yielded accurate results at a 95% confidence limit. (C) 2014 Elsevier B.V. All rights reserved.

Place, publisher, year, edition, pages
Elsevier , 2015. Vol. 102
Keyword [en]
Uricase; Ferrocene; Uric acid; Biosensor; Cyclic voltammetry; Differential pulse voltammetry
National Category
Biological Sciences
URN: urn:nbn:se:liu:diva-114967DOI: 10.1016/j.bioelechem.2014.11.001ISI: 000348953600001PubMedID: 25461755OAI: diva2:794158

Funding Agencies|Department of Biotechnology, Government of West Bengal [1695/71]

Available from: 2015-03-10 Created: 2015-03-06 Last updated: 2015-03-10

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