Generation and characterization of a functional Nanobody against the vascular endothelial growth factor receptor-2; angiogenesis cell receptor

Mahdi Behdani; Sirous Zeinali; Hossein Khanahmad; Morteza Karimipour; Nader Asadzadeh; Keyhan Azadmanesh; Alireza Khabiri; Steve Schoonooghe; Mahdi Habibi Anbouhi; Gholamreza Hassanzadeh-Ghassabeh; Serge Muyldermans

doi:10.1016/j.molimm.2011.11.013

Generation and characterization of a functional Nanobody against the vascular endothelial growth factor receptor-2; angiogenesis cell receptor

Mol Immunol. 2012 Feb;50(1-2):35-41. doi: 10.1016/j.molimm.2011.11.013. Epub 2011 Dec 29.

Authors

Mahdi Behdani¹, Sirous Zeinali, Hossein Khanahmad, Morteza Karimipour, Nader Asadzadeh, Keyhan Azadmanesh, Alireza Khabiri, Steve Schoonooghe, Mahdi Habibi Anbouhi, Gholamreza Hassanzadeh-Ghassabeh, Serge Muyldermans

Affiliation

¹ Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.

PMID: 22208996
DOI: 10.1016/j.molimm.2011.11.013

Abstract

Vascular endothelial growth factor receptor-2 (VEGFR2) is an important tumor-associated receptor and blockade of the VEGF receptor signaling can lead to the inhibition of neovascularization and tumor metastasis. Nanobodies are the smallest intact antigen binding fragments derived from heavy chain-only antibodies occurring in camelids. Here, we describe the identification of a VEGFR2-specific Nanobody, named 3VGR19, from dromedaries immunized with a cell line expressing high levels of VEGFR2. We demonstrate by FACS, that 3VGR19 Nanobody specifically binds VEGFR2 on the surface of 293KDR and HUVECs cells. Furthermore, the 3VGR19 Nanobody potently inhibits formation of capillary-like structures. These data show the potential of Nanobodies for the blockade of VEGFR2 signaling and provide a basis for the development of novel cancer therapeutics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Angiogenesis Inhibitors / immunology
Angiogenesis Inhibitors / metabolism
Angiogenesis Inhibitors / pharmacology
Animals
Antibody Specificity / immunology
Camelids, New World
Cells, Cultured
Electrophoresis, Polyacrylamide Gel
Flow Cytometry
HEK293 Cells
Human Umbilical Vein Endothelial Cells / drug effects
Human Umbilical Vein Endothelial Cells / immunology
Human Umbilical Vein Endothelial Cells / physiology
Humans
Immune Sera / immunology
Kinetics
Male
Molecular Sequence Data
Neovascularization, Physiologic / drug effects
Neovascularization, Physiologic / immunology*
Protein Binding / immunology
Sequence Homology, Amino Acid
Signal Transduction / immunology*
Single-Chain Antibodies / genetics
Single-Chain Antibodies / immunology*
Single-Chain Antibodies / pharmacology
Surface Plasmon Resonance
Vascular Endothelial Growth Factor Receptor-2 / immunology*
Vascular Endothelial Growth Factor Receptor-2 / metabolism

Substances

Angiogenesis Inhibitors
Immune Sera
Single-Chain Antibodies
Vascular Endothelial Growth Factor Receptor-2