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Characterization of a new potent, in vivo neutralizing monoclonal antibody to human vascular endothelial growth factor

  • Jean-Marc Schlaeppi
  • Gerhard Siemeister
  • Karin Weindel
  • Christian Schnell
  • Jeanette Wood
ORIGINAL PAPER

Abstract

Vascular endothelial growth factor (VEGF) is an important mediator of tumor-induced angiogenesis and represents a potential target for anticancer therapy. Therefore, we prepared a panel of monoclonal antibodies (mAb) against both the VEGF121 and VEGF165 isoforms. Three of them completely neutralized the mitogenic stimulation by VEGF of human umbilical vein endothelial cells at mAb concentrations below 0.1 μg/ml. The most potent one, with a dissociation constant (Kd) of 8 pM, inhibited, in a dose-dependent manner, VEGF-induced angiogenesis in a growth factor implant model in mice. A complete inhibition of the angiogenic response was obtained by daily intraperitoneal injections of 10 μg mAb/mouse. Angiogenesis induced by basic fibroblast growth factor was not inhibited by the mAb. Epitope mapping of the mAb, performed by competitive enzyme-linked immunosorbent assay and Western blot analysis, showed that it did not bind to the reduced and denatured monomer of VEGF. Substitutions of three residues (Q87R, G88K, Q89K), located on the major surface loop β5 to β6 of VEGF, resulted in the complete loss of binding (more than 400-fold reduction). The results suggest that the mAb binds primarily to a conformation-dependent epitope on the VEGF dimeric form, encompassing one of the loop regions involved in KDR receptor binding. The mAb with its strong neutralizing properties represents a useful agent for effective blocking of VEGF-mediated tumor neovascularization.

Key words VEGF Neutralizing antibody Epitope Angiogenesis 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Jean-Marc Schlaeppi
    • 1
  • Gerhard Siemeister
    • 2
  • Karin Weindel
    • 2
  • Christian Schnell
    • 1
  • Jeanette Wood
    • 1
  1. 1.Novartis Pharmaceuticals, Core Technology and Oncology Departments, Novartis Limited, CH-4002 Basel, SwitzerlandCH
  2. 2.Institute of Molecular Medicine, Tumor Biology Center, Freiburg, GermanyDE

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