In this paper, we propose a strategy to form nanoelectrode arrays by electrochemical deposition of the Prussian blue (PB) through highly ordered porous anodic alumina (PAA) membrane. The structure and morphology of the nanoarrays were characterized by scanning electron microscopy (SEM). As the highly ordered PB arrays can behave as an ensemble of closely spaced but isolated nanoelectrodes, the nanostructured PB arrays are successfully applied to improve the analytical performances of glucose by electrocatalytic reduction enzymatically liberated H2O2. The resulting PB based nanoelectrode arrays show a wide linear calibration range over three orders of magnitude of glucose concentrations (5.0 x 10(-6) to 8.0 x 10(-3) M) and a low detection limit of 1 mu M. Moreover, the biosensor exhibits other good characteristics, such as short response time, high selectivity, excellent operation stability. In addition, effects of the glucose oxidase (GOx) loading, applied potential and pH on the biosensor performance were also discussed. (c) 2006 Elsevier B.V. All rights reserved.