Modeling the response function of dual-enzyme microbiosensors.

A general theoretical model for competitive dual-enzyme microbiosensors based on self-assembled monolayers (SAM) is presented. The model is derived for amperometric dual-enzyme ATP sensors and provides excellent agreement with experimental ATP measurements at 25 microm diameter microelectrodes. In this model, the statistical probability of a glucose molecule in competition between two enzymes, glucose oxidase (GOD)/hexokinase (HEX), at the ATP sensor surface is combined with the enzymatic reaction rate. Thereby, a simple model predicting the sensor signal for varying surface concentrations of GOD and HEX, glucose concentration, and ATP concentration is obtained. Excellent agreement of the predicted current signal with experimentally obtained sensor signals was achieved at ATP concentrations between 10 and 300 microM in a buffer containing glucose at physiologically relevant levels. Consequently, the development time for new dual-enzyme biosensors can be reduced, and an analytical model for the sensor response function is provided facilitating the calibration of enzymatic biosensors.