An electrochemical biosensor for determination of hydrogen peroxide (H2O2)was fabricated, based on the electrostatic immobilization of horseradish peroxidase (HRP)with one-dimensional gold nanowires (Au NWs)and TiO2 nanoparticles (nano-TiO2)on a gold electrode. The nano-TiO2 can give a biocompatible microenvironment and compact film, and the Au NWs can provide fast electron transferring rate and greatly add the amount of HRP molecules immobilized on the electrode surface. Au NWs were characterized by ultraviolet-visible spectra and transmission electron microscope. The electrode modification process was probed by cyclic voltammetry and electrochemical impedance spectroscopy. Chronoamperometry was used to study the electrochemical performance of the resulting biosensor. Under optimal conditions, the linear range for the determination of H2O2 was from 2.3 9 10-6 to 2.4 9 10-3 M with a detection limit of 7.0 9 10-7 M (S/N = 3). Moreover, the proposed biosensor showed superior stability and high sensitivity.