A recombinant, fully human, bispecific antibody neutralizes the biological activities mediated by both vascular endothelial growth factor receptors 2 and 3

Mol Cancer Ther. 2005 Mar;4(3):427-34. doi: 10.1158/1535-7163.MCT-04-0261.

Abstract

Vascular endothelial growth factors (VEGF) and their receptors (VEGFR) have been implicated to play important roles in tumor-associated angiogenesis and lymphangiogenesis, and hence in tumor growth and metastasis. We previously produced a number of fully human antibodies directed against VEGF receptor 2 (VEGFR2) and VEGF receptor 3 (VEGFR3) and showed that these antibodies are capable of inhibiting growth factor (VEGF and VEGF-C)-induced receptor activation, migration, and proliferation of human endothelial cells. In this report, we constructed and produced a bispecific antibody, a diabody, using the variable domain genes of two neutralizing antibodies, IMC-1121 to VEGFR2 and hF4-3C5 to VEGFR3. The diabody binds to both VEGFR2 and VEGFR3 in a dose-dependent manner, and blocks interaction between VEGF/VEGFR2, VEGF-C/VEGFR2, and VEGF-C/VEGFR3. In cell-based assays, the diabody neutralized both VEGF and VEGF-C-stimulated activation of VEGFR2, VEGFR3, and p44/p42 mitogen-activated protein kinase in endothelial cells. Furthermore, the diabody was able to inhibit both VEGF and VEGF-C-induced migration of endothelial cells. Taken together, our results suggest that a dual blockade of both VEGFR2 and VEGFR3 simultaneously may represent a more potent approach to effective cancer therapy.

MeSH terms

  • Animals
  • Antibodies, Bispecific / chemistry*
  • Aorta / metabolism
  • Cattle
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Dose-Response Relationship, Immunologic
  • Endothelium, Vascular / cytology
  • Humans
  • Immunoglobulin Fragments / chemistry
  • Immunotherapy / methods*
  • Kinetics
  • MAP Kinase Signaling System
  • Neoplasms / immunology
  • Neoplasms / therapy*
  • Phosphorylation
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Umbilical Veins / cytology
  • Vascular Endothelial Growth Factor Receptor-2 / chemistry*
  • Vascular Endothelial Growth Factor Receptor-2 / immunology
  • Vascular Endothelial Growth Factor Receptor-3 / chemistry*
  • Vascular Endothelial Growth Factor Receptor-3 / immunology

Substances

  • Antibodies, Bispecific
  • Immunoglobulin Fragments
  • Recombinant Proteins
  • Vascular Endothelial Growth Factor Receptor-2
  • Vascular Endothelial Growth Factor Receptor-3