In this paper, we fabricated a bionanocomposite film of glucose oxidase/Pt nanoparticles/graphene-chitosan (GOD/PtNPs/GR-Chit) for glucose sensing. The hybrid bionanocomposites modified GCE were characterized by scanning electron microscopy (SEM), cyclic voltammetry, and amperometric i-t curve. It was found that the PtNPs were uniformly deposited on the surface of GR-Chit hybrid film. The resultant PtNPs/GR-Chit/GCE exhibited a high electrochemical catalytic ability to hydrogen peroxide (H2O2), due to the electrocatalytic synergy of GR and PtNPs. The redox behavior of the GOD/PtNPs/GR-Chit/GCE is a surface-controlled process. Finally, we obtained the amperometric response of the GOD/PtNPs/GR-Chit/GCE toward different concentration of glucose, and also achieved a sensitive glucose oxidase biosensor with a detection limit of 4.6μM glucose.