In this study, a novel signal-increase electrochemiluminescence (ECL) biosensor has been developed for the detection of glucose based on graphene quantum dot/glucose oxidase (GQD/GO) on Ti foil. The proposed GQD with excellent ECL ability is synthesized through a green one-step strategy by the electrochemical reduction of graphene oxide quantum dot. Upon the addition of glucose, GO can catalytically oxidize glucose and the direct electron transfer between the redox centre of GO and the modified electrode also has been realized, which results in the bio-generated HO for ECL signal increase in GQD and realizes the direct ECL detection of glucose. The signal-increase ECL biosensor enables glucose detection with high sensitivity reaching 5 × 10 mol l in a wide linear range from 5 × 10 to 1.5 × 10 mol l. Additionally, the fabrication process of such GQD-based ECL biosensor is also suitable to other biologically produced HO system, suggesting the possible applications in the sensitive detection of other biologically important targets (e.g. small molecules, protein, DNA and so on).