A label-free biosensor has been fabricated using a reduced graphene oxide (RGO) and anatase titania (antTiO 2 ) nanocomposite, electrophoretically deposited onto an indium tin oxide coated glass substrate. The RGO-ant-TiO 2 nanocomposite has been functionalized with protein (horseradish peroxidase) conjugated antibodies for the specific recognition and detection of Vibrio cholerae. The presence of Ab-Vc on the RGO-ant-TiO 2 nanocomposite has been confirmed using electron microscopy, Fourier transform infrared spectroscopy and electrochemical techniques. Electrochemical studies relating to the fabricated Ab-Vc/RGO-ant-TiO 2 /ITO immunoelectrode have been conducted to investigate the binding kinetics.This immunosensor exhibits improved biosensing properties in the detection of Vibrio cholerae, with a sensitivity of 18.17 Â 10 6 F mol À1 L À1 m À2 in the detection range of 0.12-5.4 nmol L À1 , and a low detection limit of 0.12 nmol L À1 . The association (k a ), dissociation (k d ) and equilibrium rate constants have been estimated to be 0.07 nM, 0.002 nM and 0.41 nM, respectively. This Ab-Vc/RGO-ant-TiO 2 /ITO immunoelectrode could be a suitable platform for the development of compact diagnostic devices.