Rational Design and Development of Phenazine-based Electron Mediators for Enzyme-based Biosensors: Advancing Clinical Diagnostics Through Computational Exploration

Abstract

The advancement of clinical diagnostics critically depends on the development of efficient and reliable electron mediators. These mediators are pivotal in biosensors, facilitating electron transfer (ET) reactions between the redox-active centers of immobilized enzymes and a conductive electrode support, thus enhancing the sensitivity and selectivity of biosensor devices. In this study, we specifically investigate phenazine as innovative electron mediators with promising diagnostic potential. Through meticulous computational predictions, we identify candidates exhibiting not only high solubility and favorable enzyme binding energies, but also appropriate redox potentials conducive to efficient electron transfer processes. Subsequently, these selected candidates undergo rigorous validation processes to assess their performance in blood glucose test strips, demonstrating a wide linear range from 1.00mM to 32.49mM (R2=0.9976) in whole blood tests, and a strong selectivity of glucose over other interfering substances such as ascorbic acid, uric acid, xylitol and hemoglobin.