Hybrid conjugates of graphene with metallic/semiconducting nanostructures can improve the sensitivity of electrochemical sensors due to their combination of well-balanced electrical/electrocatalytic properties and superior surface-to-volume ratio. In this study, the synthesis and physical characterization of a hybrid conjugate of reduced graphene oxide and nickel nanoparticles (rGO-Ni NPs) is presented. The conjugate was further deposited onto a glassy carbon electrode as a nanocomposite film of chitosan and glucose oxidase. The electrochemical response and morphology of the films were investigated using SEM, CV, and EIS, and their applications as a glucose biosensor explored for the first time in proof-of-concept tests. The low operating potential along with the good linearity and sensitivity (up to 129 μA cm(-2) mM(-1)) found in the sub-millimolar range suggest potential applications in the self-management of hypoglycemia from blood samples or in the development of non-invasive assays for body fluids such as saliva, tears or breath.