Two-dimensional (2D) nanosheets have been extensively explored as electrode materials for the development of high-performance electrochemical biosensors due to their unique structural characteristics. Nevertheless, 2D nanosheets suffer from sheet aggregation issues limiting the electrical conductivity of layered metal sulfides or hydroxides. Here, we report high-performance glucose biosensors based on a three-dimensional (3D) aerogel composed of interconnected 2D MoS2 and graphene sheet. 3D MoS2/graphene aerogel (MGA) provides a large surface area for the effective immobilization of enzymes, and continuous framework of electrically conductive graphene sheets. Flow-injection amperometric evaluation of the glucose biosensor using a 3D MGA electrode exhibits a rapid response (similar to 4s), a linear detection range from 2 to 20 mM, a sensitivity of 3.36 mu A/mM, and a low limit of detection of 0.29 mM. Moreover, the interference response from oxidizable species, such as ascorbic acid, uric acid and dopamine is negligible at an operating potential of -0.45 V. (C) 2017 Elsevier Inc. All rights reserved.