Abstract: This study aimed to investigate the antitumor efficacy of a single-chain variable fragment JZC00 combined with 2-deoxyglucose(2-DG)on murine non-small lung cancer cell and breast cancer cell models. JZC00 was expressed by E. coli and identified using SDS-PAGE and Western blot. The combination inhibited the proliferation of LLC and 4T1 cells. The concentration of glucose and lactic acid in the medium were determined by glucose and lactate kit, respectively, then calculated the tumor cell glucose uptake inhibition rate and lactate release inhibition rate. In vivo, the tumor volume and tumor weight were analyzed after 15-day treatment. The results showed that the molecular weight of JZC00 expressed was correct, and it could inhibit the proliferation of tumor cells in vitro. JZC00 and 2-DG could inhibit the glycolysis of tumor cells, respectively, and JZC00 combined with 2-DG could inhibit glycolysis synergistically. When hypoxic microenvironment was induced in vitro, the inhibition of glycolysis by JZC00 treatment decreased. However, it was reversed with the addition of 2-DG. The in vivo models the combination showed a significantly improved tumor suppressive effect compared with JZC00 treated group, suggesting that 2-DG could improve the anti-tumor effect of anti-angiogenic antibodies and its combination has the potentialial value in the treatment of solid tumors.