Journal
BIOSENSORS & BIOELECTRONICS
Volume 19, Issue 10, Pages 1301-1307Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2003.11.023
Keywords
ATP biosensor; hexokinase; glucose oxidase; microelectrode; amperometric sensor; polymerization
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A dual enzyme electrode for the detection of adenosine-5'-triphosphate (ATP) at physiologically relevant pH levels was developed by co-immobilization of the enzymes glucose oxidase (GOD) and hexokinase (HEX) using pH-shift induced deposition of enzyme containing polymer films. Application of a simple electrochemical procedure for the co-immobilization of the enzymes at electrode surfaces exhibits a major improvement of sensitivity, response time, reproducibility, and ease of fabrication of ATP bbiosensors. Competition between glucose oxidase and hexokinase for the substrate glucose involving ATP as a co-substrate allows the determination of ATP concentrations. Notable control on the immobilization process enables fabrication of micro biosensors with a diameter of 25 mum. The presented concept provides the technological basis for a new generation of fast responding, sensitive, and robust biosensors for the detection of ATP at physiological pH values with a detection limit of 10 nmol1(-1). (C) 2003 Elsevier B.V. All rights reserved.
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