Self-encapsulation of oxidases as a basic approach to tune the upper detection limit of amperometric biosensors

This study describes a new, basic procedure for the tuning of some analytical parameters of enzymatic biosensors that are based on hydrogen peroxide-producing oxido-reductases. An amperometric biosensor based on glucose oxidase (GOx) (EC 1.1.3.4) from Penicillum vitale, immobilized on a carbon rod electrode by cross-linking with glutaraldehyde, was exploited as a model system for demonstration of the approach described here. Such an important analytical parameter as the upper detection limit was dramatically changed by the formation of a polypyrrole conducting polymer layer by the GOx-induced polymerization of polypyrrole (Ppy). An increase in the upper detection limits for differently modified electrodes was estimated by calculation of the apparent Michaelis-Menten constant [K(M(app))]. A significant increase in the long-term stability of the GOx-based electrode modified by Ppy (GOx/Ppy) was detected compared with that of an unmodified one. Further application of this approach, based on the self-encapsulation of glucose oxidase and other oxidases, is predicted for such biosensors where extension of the detection rate as well as K(M(app)) are required.