Isolation and characterization of neutralizing monoclonal antibodies to human vascular endothelial growth factor/vascular permeability factor121 (VEGF/VPF121).
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.
Asano
M
M
Bioscience Research Department, Tsukuba Research Laboratory, Toagosei Co. Ltd., Ibaraki, Japan.
Yukita
A
A
Matsumoto
T
T
Matsuo
K
K
Kondo
S
S
Suzuki
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000628:therapy; D016228:Endothelial Growth Factors / Q000037:antagonists & inhibitors* / Q000276:immunology*; D005260:Female; D006131:Growth Inhibitors / Q000494:pharmacology; D006801:Humans; D006825:Hybridomas; D008113:Liver Neoplasms / Q000473:pathology / Q000628:therapy; D008222:Lymphokines / Q000037:antagonists & inhibitors* / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000911:Antibodies, Monoclonal; D016228:Endothelial Growth Factors; D006131:Growth Inhibitors; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors