Angiogenesis mediated by vascular endothelial growth factor (VEGF) is a hallmark of glioblastoma. Based on the response rate and improved progression-free survival， although not on overall survival， the 149-kDa anti-VEGF-A IgG antibody bevacizumab (Avastin) has been approved in the United States and Japan for recurrent glioblastoma and in Japan for newly diagnosed glioblastoma; however， it is not approved in the EU. Here we characterize the biologic activity of DLX1008， a 26-kDa anti-VEGF-A single-chain antibody fragment that shows 30-fold stronger affinity to human VEGF-A than bevacizumab. The small molecular size of DLX1008 is predicted to result in improved target coverage over bevacizumab. DLX1008 showed superiority to bevacizumab in the inhibition of VEGF-A binding to VEGF receptor (VEGFR) 1 in enzyme-linked immunosorbent assay by a factor of around 10 and comparable efficacy for the inhibition of VEGF-A-stimulated VEGFR2 dimerization. In a tube-formation assay with human cerebral microvascular endothelial cells， DLX1008 was at least as active as bevacizumab. In vivo， DLX1008 delayed growth in a mouse subcutaneous U87 xenograft model ( = 0.0021) and improved survival in a mouse orthotopic U87 xenograft model ( = 0.00026). Given the exceptionally high affinity and small molecular size of DLX1008， these data warrant further clinical development of DLX1008 as an antiangiogenic agent in glioblastoma.