Molecular or ion transport through a single nanopore or nanopore array is a key process in a number of applications including: molecular separation, biosensing, energy storage, nanofluidics and nanoelectronics. In this study, the electrochemical and electrical properties of nanopore arrays were investigated by electrochemical impedance spectroscopy (EIS), to explore their capability towards the development of improved nanopore biosensing devices. Anodic aluminum oxide (AAO) nanopore arrays with square patterns were fabricated by a photolithographic technique for use as a biosensing platform. To demonstrate the potential capability of these nanoporous arrays for biosensing applications, their internal surfaces were functionalized with a self-assembled monolayer of phosphonic acid; followed by covalent bonding with the model binding molecule streptavidin. The sensing performance of the prepared sensing device was characterized by EIS using different concentrations of biotin as probe molecule. The changes in impedance signal in response to the specific binding of biotin molecules to the streptavidin inside nanopores were monitored and explored depending on pore morphology. The results obtained for this system exhibit a high level of sensitivity and selectivity, which could provide the basis for the development of superior biosensing devices. ; K. Kant, M. Kurkuri, J. Yu, J. G. Shapter, C. Priest, D. Losic