Open AccessMonoclonal antibodies to vascular endothelial growth factor‐D block its interactions with both VEGF receptor‐2 and VEGF receptor‐3
Author(s) -
Achen Marc G.,
Roufail Sally,
Domagala Teresa,
Catimel Bruno,
Nice Edouard C.,
Geleick Detlef M.,
Murphy Roger,
Scott Andrew M.,
Caesar Carol,
Makinen Taija,
Alitalo Kari,
Stacker Steven A.
Publication year - 2000
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.2000.01257.x
Subject(s) - vascular endothelial growth factor , receptor , angiogenesis , biology , kinase insert domain receptor , receptor tyrosine kinase , microbiology and biotechnology , vascular endothelial growth factor c , vascular endothelial growth factor a , chemistry , cancer research , vegf receptors , biochemistry
Vascular endothelial growth factor‐D (VEGF‐D), the most recently discovered mammalian member of the VEGF family, is an angiogenic protein that activates VEGF receptor‐2 (VEGFR‐2/Flk1/KDR) and VEGFR‐3 (Flt4). These receptor tyrosine kinases, localized on vascular and lymphatic endothelial cells, signal for angiogenesis and lymphangiogenesis. VEGF‐D consists of a central receptor‐binding VEGF homology domain (VHD) and N‐terminal and C‐terminal propeptides that are cleaved from the VHD to generate a mature, bioactive form consisting of dimers of the VHD. Here we report characterization of mAbs raised to the VHD of human VEGF‐D in order to generate VEGF‐D antagonists. The mAbs bind the fully processed VHD with high affinity and also bind unprocessed VEGF‐D. We demonstrate, using bioassays for the binding and cross‐linking of VEGFR‐2 and VEGFR‐3 and biosensor analysis with immobilized receptors, that one of the mAbs, designated VD1, is able to compete potently with mature VEGF‐D for binding to both VEGFR‐2 and VEGFR‐3 for binding to mature VEGF‐D. This indicates that the binding epitopes on VEGF‐D for these two receptors may be in close proximity. Furthermore, VD1 blocks the mitogenic response of human microvascular endothelial cells to VEGF‐D. The anti‐(VEGF‐D) mAbs raised to the bioactive region of this growth factor will be powerful tools for analysis of the biological functions of VEGF‐D.