Isolation and Characterization of Neutralizing Monoclonal Antibodies to Human Vascular Endothelial Growth Factor/Vascular Permeability Factor₁₂₁(VEGF/VPF₁₂₁)

Abstract

We have established monoclonal antibodies (MAbs) against human vascular endothelial growth factor/vascular permeability factor121 (VEGF/VPF121). Two (MV101 and MV303) of the 28 MAbs neutralized the mitogenic activity of VEGF/VPF121 on human umbilical vein endothelial cells (HUVEC) in a dose-dependent manner. Both of the MAbs reacted to VEGF/VPF121 and also VEGF/VPF165 with somewhat different binding properties in a sandwich-type enzyme-linked immunosorbent assay (ELSIA). The binding of MV101 and MV303 to VEGF/VPF121 was competitive, but MV415, another anti-VEGF/VPF121 MAb without neutralizing activity, did not complete with either of the antibodies. MV101 and MV303 specifically recognized the native form of VEGF/VPF121 and VEGF/VPF165 in Western blotting. They did not react with VEGF/VPF when the antigens were fractionated under reducing conditions. These observations suggested that MV101 and MV303 might recognize the epitopes closely located on the configuration of VEGF/VPF121 molecule and the epitopes recognized by MV101 and MV303 may play an important role in the VEGF/VPF-receptor signal transduction. These MAbs significantly suppressed the growth of a human hepatoma, PLC/PRF/5, in vivo.

Growth inhibition of astrocytoma cells by farnesyl transferase inhibitors is mediated by a combination of anti-proliferative, pro-apoptotic and anti-angiogenic effects

“…We thus investigated whether this enhanced in vivo e ect may relate to an additional anti-angiogenic e ect of the agent. Evidence from our laboratory and from others has demonstrated the importance of the angiogenic factor Vascular Endothelial Growth Factor (VEGF) in the pathogenesis of high grade astrocytic tumours (Asano et al, 1995;Feldkamp et al, 1999;Melnyk et al, 1996;Millauer et al, 1996;Saleh et al, 1996). Astrocytomas secrete high levels of the potent and endothelial speci®c angiogenic factor VEGF (Plate et al, 1992), whose expression through increased transcription or enhanced mRNA stability is highly regulated by hypoxia Goldberg and Schneider, 1994;Levy et al, 1996).…”

Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets: significance for VEGF measurements and cancer biology

“…Use of neutralizing antibodies to VEGF or transfection with antisense VEGF cDNA have been shown to result in inhibition of growth of tumour cell lines in nude mice and inhibition of metastatic spread (Asano et al, 1995;Warren et al, 1995;Claffey et al, 1996;Saleh et al, 1996). Serum concentrations of VEGF have been examined in patients with a range of cancers, with higher serum levels of VEGF being apparent in some patients (Kondo et al, 1994;Ferrari and Scagliotti, 1996;Takano et al, 1996;Dirix et al, 1997).…”

Effect of VEGF receptor inhibitor PTK787/ZK222548 combined with ionizing radiation on endothelial cells and tumour growth

“…Evidence for the importance of VEGFinduced angiogenesis in tumour growth includes the observation that neutralizing antibodies against VEGF or dominant negative soluble receptors inhibit the growth of primary and metastatic experimental tumours (Kim et al, 1993;Asano et al, 1995;Borgestrom et al, 1996;Goldman et al, 1998;Kanai et al, 1998).…”