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Characterisation of horseradish peroxidase immobilisation on an electrochemical biosensor by colorimetric and amperometric techniques

Morrin, Aoife and Guzman, Alicia and Killard, Anthony J. and Pingarron, Jose M. and Smyth, Malcolm R. (2003) Characterisation of horseradish peroxidase immobilisation on an electrochemical biosensor by colorimetric and amperometric techniques. Biosensors and Bioelectronics, 18 (5-6). pp. 715-720. ISSN 0956-5663

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Abstract

This study presents the use of complementary colorimetric and amperometric techniques to measure the quantity of protein or enzyme immobilised onto a carbon paste electrode modified with a layer of electrodeposited polyaniline. By applying a solution of bovine serum albumin at 0.75 mg/ml, efficient blocking of the electrode from electroactive species in the bulk solution could be achieved. When the horseradish peroxidase was immobilised on the electrode, optimal amperometric responses from hydrogen peroxide reduction were achieved at approximately the same concentration. The mass of enzyme immobilised at this solution concentration was determined by a colorimetric enzyme assay to be equivalent to the formation of a protein monolayer. Under these conditions, amperometric responses from the immobilised layer are maximised and non-specific bulk solution interactions are minimised. At higher immobilised protein concentrations, diminished amperometric responses may be due to inhibited diffusion of hydrogen peroxide to enzyme which is in electronic communication with the electrode surface, or impeded electron transfer.

Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:horseradish peroxidase; enzyme immobilisation; screen-printed electrode; polyaniline; amperometry; colorimetric enzyme assay;
Subjects:Biological Sciences > Enzymology
Physical Sciences > Chemistry
Biological Sciences > Biosensors
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Chemical Sciences
Research Initiatives and Centres > National Centre for Sensor Research (NCSR)
Publisher:Elsevier
Official URL:http://dx.doi.org/10.1016/S0956-5663(03)00003-4
ID Code:54
Deposited On:08 Nov 2006 by DORAS Administrator. Last Modified 30 Jan 2009 12:01

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