Multilayered assembly of dendrimers with enzymes on gold: Thickness-controlled biosensing interface

A new approach to construct a multilayered enzyme film on the Au surface for use as a biosensing interface is described, The film was prepared by alternate layer-by-layer depositions of G4 poly(amidoamine) dendrimers and periodate-oxidized glucose oxidase (GOx), The cyclic voltammograms obtained from the Au electrodes modified with the GOx/dendrimer multilayers revealed that bioelectrocatalytic response is directly correlated to the number of deposited bilayers, that is, to the amount of active enzyme immobilized on the Au electrode surface. From the analysis of voltammetric signals, the coverage of active enzyme per GOx/dendrimer bilayer during the multilayer-forming steps was estimated, which demonstrates that the multilayer is constructed in a spatially ordered manner. Also, with the ellipsometric measurements, a linear increment of the film thickness was registered, supporting the formation of the proposed multilayered structure. The E5D5 electrode showed the sensitivity of 14.7 mu A.mM(-1) glucose.cm(-2) and remained stable over 20 days under day-by-day calibrations. The proposed method is simple and would be applicable to the constructions of thickness- and sensitivity-controllable biosensing interfaces composed of multienzymes as well as a single enzyme.