A simple strategy for one-step construction of bienzyme biosensor by in-situ formation of biocomposite film through electrodeposition

One-step construction of horseradish peroxidase (HRP) and glucose oxidase (GOD) bienzyme biosensor was established based on a simple and controllable electrodeposition approach by in-situ formation of biocomposite film on electrode. With hydrogen peroxide (H2O2) instantly generated by GOD-catalyzed oxidation of glucose, quantitative measurement of glucose could be achieved. In the proposed electrodeposition strategy, Concanavalin A (Con A) was applied as bifunctional cross-linker for in-situ incorporating of HRP–GOD in biocompatible chitosan (CS) matrix due to its biospecific affinity interaction with CS and glycoproteins. Scanning electron microscopy showed that the HRP–GOD/Con A/CS biocomposite film possessed highly porous surface. The developed bienzyme biosensor exhibited a fast amperometric response for the determination of glucose. The linear response of the developed biosensor for the determination of glucose ranged from 1.0 × 10−6 to 2.2 × 10−4 M with a detection limit of 6.7 × 10−7 M. The biosensor can be used to determine glucose in serum directly. The biosensor presented high stability owing to the design of the introduction of Con A and instant generation of H2O2 with bienzyme system. The one-step construction of biosensor based on non-manual technique in biocompatible environment was direct and facile.