Graphene/Fe 3 O 4 nanocomposite was prepared for the immobilization of hemoglobin (Hb) to improve the electron transfer between Hb and glass carbon electrode (GCE). The characterization of nanocomposites was described by transmission electron microscopy, Fourier transform infrared, Raman spectroscopy, and X-ray photoelectron spectroscopy, respectively. The electrochemistry of Hb on the graphene/ Fe 3 O 4 -based GCE was investigated by cyclic voltammetry and amperometric measurement. The modified electrode showed a wide linear range from 0.25 μmol/L to 1.7 mmol/ L with a correlation coefficient of 0.9967. The detection limit of the H 2 O 2 biosensor was estimated at 6.0×10 −6 mol/L at a signal-to-noise ratio of 3.