Laser-Induced Graphene Arrays-Based Three-Phase Interface Enzyme Electrode for Reliable Bioassays.

Electrochemical oxidase biosensors have been widely applied in healthcare, environmental measurements and the biomedical field. However, the low and fluctuant oxygen levels in solution and the high anodic detection potentially restrict the assay accuracy. To address these problems, in this work, we constructed a three-phase interface enzyme electrode by sequentially immobilizing H ₂ O ₂ electrocatalysts and an oxidase layer on a superhydrophobic laser-induced graphene (LIG) array substrate. The LIG-based enzyme electrode possesses a solid-liquid-air three-phase interface where constant and sufficient oxygen can be supplied from the air phase to the enzymatic reaction zone, which enhances and stabilizes the oxidase kinetics. We discovered that the enzymatic reaction rate is 21.2-fold improved over that of a solid-liquid diphase system where oxygen is supplied from the liquid phase, leading to a 60-times wider linear detection range. Moreover, the three-phase enzyme electrode can employ a cathodic measuring principle for oxidase catalytic product H ₂ O ₂ detection, which could minimize interferences arising from oxidizable molecules in biofluids and increase the detection selectivity. This work provides a simple and promising approach to the design and construction of high-performance bioassay systems.