title: Inhibition of angiogenesis in human endothelial cell using VEGF specific nanobody.
abstract: Angiogenesis is an important step in tumor development and metastasis. Vascular endothelial growth factor (VEGF) plays an important role in progression of angiogenesis. VEGF121 and VEGF165 are the most relative forms of VEGF family which contain the full biological activity. Nanobodies derived from camelidae are the smallest biding site of antigen. Unique characteristic of nanobodies make them as a useful candidate for research. In this report, we describe the isolation of VEGF specific nanobodies from dromedaries immunized with purified VEGF antigen using phage display. Four clones that showed the highest signal value in ELISA experiment were selected and expressed as a His-tagged fusion protein. Four selected nanobodies were reacted strongly to VEGF in cross-reactivity assay. The binding affinity of selected nanobodies named Nb22, Nb23, Nb35 and Nb42 were differed from 0.1 to 60nM. The nanobodies inhibited endothelial cell proliferation or tube formation in response of VEGF in a dose-dependent manner. These results indicate the potential of nanobodies in inhibition of VEGF and represent a promising candidate for cancer research and therapeutics.
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Fatemeh
authors: F
authors: Kazemi-Lomedasht
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Mahdi
authors: M
authors: Behdani
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Kamran Pooshang
authors: KP
authors: Bagheri
authors: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors: Mahdi
authors: M
authors: Habibi-Anbouhi
authors: Immunology Department, Hybridoma Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Mohsen
authors: M
authors: Abolhassani
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Roghaye
authors: R
authors: Arezumand
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors: Delavar
authors: D
authors: Shahbazzadeh
authors: Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran. Electronic address: mirzahoseini@pasteur.ac.ir.
authors: Hasan
authors: H
authors: Mirzahoseini
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D002162:Camelus / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000276:immunology*; D004730:Endothelium, Vascular / Q000276:immunology; D006801:Humans; D009389:Neovascularization, Pathologic / Q000276:immunology*; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D061905:Single-Domain Antibodies / Q000276:immunology*; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology*
chemical_list: D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Angiogenesis; Nanobody; Phage display; VEGF