title: Development of a Novel Human scFv Against EGFR L2 Domain by Phage Display Technology.
abstract: Epidermal growth factor receptor (EGFR) as a transmembrane tyrosine kinase receptor frequently overexpresses in tumors with epithelial origin. The L2 domain from extracellular part of EGFR is involved in ligand binding and the blockage of this domain prevents activation of related signaling pathways. This study was aimed to develop a novel human scFv against EGFR L2 domain as a promising target for cancer therapy. The L2 recombinant protein was purified and used for panning a human scFv phage library (Tomlinson I). In this study, a novel screening strategy was applied to select clones with high binding and enrichment of rare specific phage clones of the L2 protein. After five biopanning rounds several specific clones were isolated which among them one phage clone with high binding was purified for further analysis. The specific interaction of selected clone against target antigen was confirmed by ELISA and western blotting. Immunofluorescence staining showed that purified scFv binds to A431 cells surface, displaying EGFR surface receptor. In the present study, we isolated for the first time a novel human scFv against EGFR L2 domain. This study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFR overexpressing cancers using this novel human anti-L2 ScFv.
authors: Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz. Iran.
authors: Leila
authors: L
authors: Rahbarnia
authors: Daneshgah Street, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz. Iran.
authors: Safar
authors: S
authors: Farajnia
authors: Drug Applied Research Center, Tabriz University of Medical Sciences, Daneshgah Street, Tabriz. Iran.
authors: Hossein
authors: H
authors: Babaei
authors: Immunology Research Center, Tabriz University of Medical Sciences, Daneshgah Street, Tabriz. Iran.
authors: Jafar
authors: J
authors: Majidi
authors: Biotechnology Department, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Science. 0.
authors: Kamal
authors: K
authors: Veisi
authors: Drug Applied Research Center, Tabriz University of Medical Sciences, Daneshgah Street, Tabriz. Iran.
authors: Shiva Ahdi
authors: SA
authors: Khosroshahi
authors: Maragheh University of Medical Sciences, Maragheh. Iran.
authors: Asghar
authors: A
authors: Tanomand
mesh_terms: D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D009369:Neoplasms / Q000188:drug therapy*; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000032:analysis* / Q000276:immunology / Q000494:pharmacology*
chemical_list: D019151:Peptide Library; D057127:Single-Chain Antibodies; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: EGFR L2 domain; Human single chain antibody; cancer; phage display