title: Quantifying protein adsorption and function at nanostructured materials: enzymatic activity of glucose oxidase at GLAD structured electrodes.
abstract: Nanostructured materials strongly modulate the behavior of adsorbed proteins; however, the characterization of such interactions is challenging. Here we present a novel method combining protein adsorption studies at nanostructured quartz crystal microbalance sensor surfaces (QCM-D) with optical (surface plasmon resonance SPR) and electrochemical methods (cyclic voltammetry CV) allowing quantification of both bound protein amount and activity. The redox enzyme glucose oxidase is studied as a model system to explore alterations in protein functional behavior caused by adsorption onto flat and nanostructured surfaces. This enzyme and such materials interactions are relevant for biosensor applications. Novel nanostructured gold electrode surfaces with controlled curvature were fabricated using colloidal lithography and glancing angle deposition (GLAD). The adsorption of enzyme to nanostructured interfaces was found to be significantly larger compared to flat interfaces even after normalization for the increased surface area, and no substantial desorption was observed within 24 h. A decreased enzymatic activity was observed over the same period of time, which indicates a slow conformational change of the adsorbed enzyme induced by the materials interface. Additionally, we make use of inherent localized surface plasmon resonances in these nanostructured materials to directly quantify the protein binding. We hereby demonstrate a QCM-D-based methodology to quantify protein binding at complex nanostructured materials. Our approach allows label free quantification of protein binding at nanostructured interfaces.
authors: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark.
authors: Uffe B
authors: UB
authors: Jensen
authors: Elena E
authors: EE
authors: Ferapontova
authors: Duncan S
authors: DS
authors: Sutherland
mesh_terms: D000327:Adsorption; D015374:Biosensing Techniques; D004566:Electrodes; D005949:Glucose Oxidase / Q000737:chemistry* / Q000378:metabolism*; D049329:Nanostructures / Q000737:chemistry*; D011485:Protein Binding; D013499:Surface Properties
chemical_list: D005949:Glucose Oxidase