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
x
) 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
x
can catalytically oxidize glucose and the direct electron transfer between the redox centre of GO
x
and the modified electrode also has been realized, which results in the bio-generated H
2
O
2
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
−6
mol l
−1
in a wide linear range from 5 × 10
−6
to 1.5 × 10
−3
mol l
−1
. Additionally, the fabrication process of such GQD-based ECL biosensor is also suitable to other biologically produced H
2
O
2
system, suggesting the possible applications in the sensitive detection of other biologically important targets (e.g. small molecules, protein, DNA and so on).