title: Isolation and characterization of anti c-met single chain fragment variable (scFv) antibodies.
abstract: The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques. Three c-Met specific scFv (ES1, ES2, and ES3) were selected following five rounds of panning procedures. The scFv showed specific binding to c-Met receptor, and significantly inhibited proliferation responses of a human colorectal carcinoma cell line (HCT-116). Moreover, anti- apoptotic effects of selected scFv antibodies on the HCT-116 cell line were also evaluated using Annexin V/PI assays. The results demonstrated rates of apoptotic cell death of 46.0, 25.5, and 37.8% among these cells were induced by use of ES1, ES2, and ES3, respectively. The results demonstrated ability to successfully isolate/char-acterize specific c-Met scFv that could ultimately have a great therapeutic potential in immuno-therapies against (colorectal) cancers.
authors.affiliation: a Immunology Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.|b Department of Immunology , Tabriz University of Medical Sciences , Tabriz , Iran.|c Immunology Department, Hybridoma Laboratory , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Elmira Safaie
authors.initials: ES
authors.lastname: Qamsari
authors.affiliation: c Immunology Department, Hybridoma Laboratory , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Sharifzadeh
authors.affiliation: a Immunology Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.|b Department of Immunology , Tabriz University of Medical Sciences , Tabriz , Iran.|c Immunology Department, Hybridoma Laboratory , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Salman
authors.initials: S
authors.lastname: Bagheri
authors.affiliation: d Department of Immunology , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Farhad
authors.initials: F
authors.lastname: Riazi-Rad
authors.affiliation: e Monoclonal Anti-body Research Center , Avicenna Research Institute , Tehran , Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Younesi
authors.affiliation: c Immunology Department, Hybridoma Laboratory , Pasteur Institute of Iran , Tehran , Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Abolhassani
authors.affiliation: f Department of Biotechnology, Pharmaceutical Sciences Branch , Islamic Azad University , Tehran , Iran.
authors.forename: Sepideh Safaei
authors.initials: SS
authors.lastname: Ghaderi
authors.affiliation: a Immunology Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Behzad
authors.initials: B
authors.lastname: Baradaran
authors.affiliation: g Liver and Gastrointestinal Diseases Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Somi
authors.affiliation: g Liver and Gastrointestinal Diseases Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Yousefi
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D063646:Carcinogenesis; D048708:Cell Growth Processes / Q000187:drug effects; D061505:Cell Surface Display Techniques; D015179:Colorectal Neoplasms / Q000276:immunology / Q000378:metabolism* / Q000628:therapy*; D045325:HCT116 Cells; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D057127:Single-Chain Antibodies; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met
keywords: HGF; c-Met; colorectal cancer; phage display; scFv