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.