Identification of the Residues in the Extracellular Region of KDR Important for Interaction with Vascular Endothelial Growth Factor and Neutralizing Anti-KDR Antibodies

Abstract

The kinase domain receptor (KDR) of vascular endothelial growth factor (VEGF) is the main human receptor responsible for the angiogenic activity of VEGF. The extracellular region of KDR is comprised of seven immunoglobulin-like domains, of which the first three have been shown to be required for ligand binding.

Tailoring in Vitro Selection for a Picomolar Affinity Human Antibody Directed against Vascular Endothelial Growth Factor Receptor 2 for Enhanced Neutralizing Activity

“…Porcine aortic endothelial cells transfected with KDR (PAE-KDR) was generated at ImClone Systems Incorporated (New York, NY) and maintained in Dulbecco's modified Eagle's medium containing 10% fetal calf serum. The soluble KDR-alkaline phosphatase fusion protein (KDR-AP), its extracellular Ig domain (ECD) deletion variant-AP fusions, and VEGF 165 proteins were produced as previously described (26). IMC-1C11 and MAB664, two mAb directed against KDR, were generated at ImClone Systems Incorporated as previously described (10,12,27).…”

“…IMC-1C11) or domain 3 alone (e.g. MAB664) (26,27). Both IMC-2C6 and IMC-1121 bind equally well to full-length KDR ECD, KDR(Ig1-3), and KDR(Ig3), thus locating their binding epitope(s) within the single Ig domain 3 of KDR ECD (Fig.…”

Inhibition of both paracrine and autocrine VEGF/ VEGFR-2 signaling pathways is essential to induce long-term remission of xenotransplanted human leukemias

“…E merging data suggests that targeting tumor angiogenesis with agents that block endothelial proliferation, such as with neutralizing vascular endothelial growth factor receptor (VEGFR)-specific antibodies, results in delayed growth of solid tumor cell lines implanted into mice (1). However, the exact mechanism whereby inhibition of the VEGF-VEGFR axis results in the regression of tumor tissue is not well studied.…”

Single Variable Domain-IgG Fusion

“…VEGF protein was expressed in baculovirus and purified at ImClone Systems (New York) following the procedure described elsewhere (22). eEnd.1 cells, a murine endothelium line transformed by polyoma middle T oncogene, was a kind gift from Dr. M. Pepper of University Medical Center, Geneva, Switzerland.…”