Adsorption of Preformed Microgel–Enzyme Complexes as a Novel Strategy toward Engineering Microgel-Based Enzymatic Biosensors

Abstract

A novel approach to surface modification, which consists of the adsorption of microgel–enzyme complexes preformed in solution, is highlighted. Accordingly, the microgel–enzyme complexes were formed due to the electrostatic interaction of the oppositely charged interacting components, that is, a cationic poly(N-isopropylacrylamide)-based microgel and glucose oxidase taken as a model enzyme. The spontaneous adsorption of the prepared microgel–enzyme complexes, examined by means of quartz crystal microbalance with dissipation monitoring and atomic force microscopy, was observed, resulting in the formation of well-adhered microgel–enzyme coatings. Further, the preformed microgel–enzyme complexes were adsorbed onto the modified graphite-based screen-printed electrodes, and their enzymatic responses were determined by means of amperometry, demonstrating a remarkable analytical performance toward the quantification of β-D-glucose in terms of high sensitivity (0.0162 A × M−1 × cm−2), a low limit of detection (1 μM), and an expanded linear range (1–2000 μM). The fabricated microgel–enzyme biosensor constructs were found to be very stable against manifold-repeated measurements. Finally, the pH- or salt-induced release of glucose oxidase from the adsorbed preformed microgel–enzyme complexes was demonstrated. The findings obtained for the microgel–enzyme coatings prepared via adsorption of the preformed microgel–enzyme complexes were compared to those found for the microgel–enzyme coatings fabricated via a previously exploited two-stage sequential adsorption, which includes the adsorption of the microgel first, followed by the electrostatic binding of glucose oxidase by the adsorbed microgel.