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Enzyme Immobilization Strategies and Electropolymerization Conditions to Control Sensitivity and Selectivity Parameters of a Polymer-Enzyme Composite Glucose Biosensor.doc

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Enzyme Immobilization Strategies and Electropolymerization Conditions to Control Sensitivity and Selectivity Parameters of a Polymer-Enzyme Composite Glucose Biosensor

Sensors 2010, 10, 6439-6462; doi:10.3390/s100706439 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Article Enzyme Immobilization Strategies and Electropolymerization Conditions to Control Sensitivity and Selectivity Parameters of a Polymer-Enzyme Composite Glucose Biosensor Sharon A. Rothwell, Sarah J. Killoran and Robert D. O’Neill * UCD School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland * Author to whom correspondence should be addressed; E-Mail: robert.oneill@ucd.ie; Tel.: +353-1-716-2314. Received: 13 April 2010; in revised form: 21 May 2010 / Accepted: 13 June 2010 / Published: 30 June 2010 Abstract: In an ongoing programme to develop characterization strategies relevant to biosensors for in-vivo monitoring, glucose biosensors were fabricated by immobilizing the enzyme glucose oxidase (GOx) on 125 μm diameter Pt cylinder wire electrodes (PtC), using three different methods: before, after or during the amperometric electrosynthesis of poly(ortho-phenylenediamine), PoPD, which also served as a permselective membrane. These electrodes were calibrated with H2O2 (the biosensor enzyme signal molecule), glucose, and the archetypal interference compound ascorbic acid (AA) to determine the relevant polymer permeabilities and the apparent Michaelis-Menten parameters for glucose. A number of selectivity parameters were used to identify the most successful design in terms of the balance between substrate sensitivity and interference blocking. For biosensors electrosynthesized in neutral buffer under the present conditions, entrapment of the GOx within the PoPD layer produced the design (PtC/PoPD-GOx) with the highest linear sensitivity to glucose (5.0 ± 0.4 μA cm?2 mM?1), good linear range (KM = 16 ± 2 mM) and response time ( 2 s), and the greatest AA blocking (99.8% for 1 mM AA). Further optimiz