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Nickel ferrite (NiFe2O4) nanoparticles have been dispersed in chitosan solution in order to fab-ricate nanocomposite films. Horseradish peroxidase (HRP) has been immobilized onto this chitosan-NiFe2O4 nanocomposite film via physical adsorption. The size of the NiFe2O4 nanoparticles has been estimated us-ing X-ray diffraction patt and scanning electron microscopy (SEM) to be 40±9 nm. The chitosan-NiFe2O4 nanocomposite film and HRP/chitosan-NiFe2O4 bioelectrode have been characterized using SEM technique. The HRP/chitosan-NiFe2O4 nanocomposite bioelectrode has a response time of 4 s, linearity as 0.3 to 12 mM of H2O2, sensitivity as 22 nA/mM. The effects of pH and the temperature of the immobilized HRP electrode have also been studied.