title: Construction and expression of an anti-VEGFR2 Nanobody-Fc fusionbody in NS0 host cell.
abstract: Angiogenesis is the formation of new blood vessels which is involved in migration, growth and differentiation of endothelial cells. This process regularly occurs during growth and development in children however, in adults is usually part of a disease process such as cancer. The vascular endothelial growth factor (VEGF) is a vital player in the vascular development and angiogenesis in physiological and pathological processes. Camelid's immune system has unique antibodies which are composed of only a heavy chain homodimer and the variable domain (VHH, Nanobody). Nanobodies are small, around 15 kDa and stable. In this study, we engineered and constructed a new Nanobody-Fc fusion protein (fusionbody) composed of an anti-VEGFR2 Nanobody and an Fc fragment of human IgG1 antibody. The recombinant vector was transfected into NS0 host cells. Stable producer clones were developed and the recombinant fusionbody was expressed and purified. Functional assay showed the anti-VEGFR2 fusionbody could bind to VEGFR2 on cell surface via VHH part and could mediate killing the targeted cells through direct cell death and complement-dependent cytotoxicity (CDC).
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran; Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Qasemi
authors.affiliation: Biotechnology Research Center, Venom and Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Behdani
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Shokrgozar
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Molla-Kazemiha
authors.affiliation: Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran. Electronic address: kouchesfehani@yahoo.com.
authors.forename: Homa
authors.initials: H
authors.lastname: Mohseni-Kuchesfahani
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran. Electronic address: habibi_m@pasteur.ac.ir.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Habibi-Anbouhi
mesh_terms: D000818:Animals; D016923:Cell Death; D002460:Cell Line; D003001:Cloning, Molecular; D003165:Complement System Proteins / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000276:immunology*; D051379:Mice; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D061905:Single-Domain Antibodies / Q000235:genetics / Q000276:immunology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000276:immunology*
chemical_list: D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; D003165:Complement System Proteins; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; Cancer; Nanobody-Fc fusion protein (fusionbody); VEGFR2