RESUMEN
Nanoporous surfaces are promising for label-free electrochemical biosensing. We formed nanopores directly on the electrode surface by means of assembling a dense layer of nonconductive nanoparticles. In our model affinity biosensor, covalent attachment of albumin protein on top of 40 nm polystyrene nanoparticles represented a capture of an analyte, resulting in blockage of the nanopores. Different bulk concentrations of the ferro/ferricyanide redox pair were probed by Faradaic electrochemical impedance spectroscopy and fast chronoamperometry. The character of the redox probe permeation towards the electrode surface differed in dependence on its concentration. These data were compared with the theoretical behavior of the free diffusion according to the Cottrell equation. Both the bulk concentration of the redox probe and the timescale of the experiment affected the performance of the electrochemical detection, demonstrating the importance of controlling these parameters in immunosensing applications.
Asunto(s)
Técnicas Biosensibles , Nanoporos , Técnicas Biosensibles/métodos , Espectroscopía Dieléctrica/métodos , Electrodos , Oxidación-Reducción , Técnicas Electroquímicas , Oro/químicaRESUMEN
Blockage of a nanopore by an analyte molecule has emerged as a promising concept for electrochemical biosensing. Nanoporous structures can be formed on the electrode surface simply by packing spherical nanoparticles in a dense planar arrangement. Modification of the nanoparticles with human serum albumin (HSA) and its interaction with the corresponding antibody (anti-HSA) can induce nanopore-blockage which significantly hinders permeation of the redox probe ([Fe(CN6)]4-/3-). Interfaces of different parameters were studied using Electrochemical Impedance Spectroscopy (EIS), and counterintuitively, the influence of charge of the nanoparticles and other immobilized entities played a substantial role in the measurement. Our study reveals dominant effects including the presence of mixed output signal and resolves corresponding EIS biosensing-related challenges. Consequently, blocking the nanopores was introduced as an efficient technique which enables the application of EIS-based biosensing to real-world analytical issues.