A smart sensing platform based on a triboelectric nanogenerator (TENG) possesses various advantages such as self-powering, convenience, real-time and biocompatibility. However, the detection limit of the TENG-based sensor is required to be improved. In this study, a high performance TENG-based glucose sensor was proposed by using the Ti3C2Tx (MXene)/graphene oxide (GO) composite electrode. The MXene and GO nanosheets are popular 2D materials which possessed high conductivity and a rich surface functional group. The MXene/GO thin films were prepared through electrostatic self-assembly technology, which can effectively impede the agglomeration of two nanoflakes. The as-prepared MXene/GO film presented outstanding mechanical property. To figure out the relationship between the nanostructure of MXene/GO film and the TENG, a series of MXene/GO-based TENG with different GO sizes was characterized. As a result, the TENG with 400 nm GO demonstrated the highest output performance. Subsequently, the optimized TENG was used in glucose detection application without the assistance of a glucose enzyme. This simple and flexible TENG shows promising potential in biosensors and non-invasive health monitoring.