Blocking the Nanopores in a Layer of Nonconductive Nanoparticles: Dominant Effects Therein and Challenges for Electrochemical Impedimetric Biosensing.

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.