Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 18 3 2019 06 08 289 299 EN Shirafkan Kordi Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran AND Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran Mohammad Rahmati-Yamchi Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran Mehdi Asghari Vostakolaei Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran Ali Etemadie Department of Medical Biotechnology, Tehran University of Medical Sciences, Tehran, Iran Abolfazl Barzegari Research Centre for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran Jalal Abdolalizadeh Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran AND Faculty of Paramedicine, Tabriz University of Medical Sciences, Tabriz, Iran 2018 08 29 2018 11 13 Vascular endothelial growth factor receptor 2 (VEGFR-2) is known as one of the important antigens playing a vital role in angiogenesis. In this study, phage display technology (PDT) was used to produce a single-chain variable fragment (scFv) antibody against a region of the domain 3 in VEGFR-2 called kinase insert domain receptor 3 (KDR3). After designing the KDR3 peptide and biopanning, a colony was chosen for scFv antibody expression. Following expression and purification; western blotting, dot blotting and immunofluorescence (IF) were used to evaluate the antibody function. Surface plasmon resonance (SPR) was also employed to measure affinity of produced antibody. Once a colony was selected and transferred to the expression host, the scFv antibody was expressed in the expected range of 28 kDa. Using a designed chromatography column, antibody purification was found to be about 95%. In this study, a novel scFv with the capability of binding to KDR3 was isolated and purified and its intracellular function was investigated and verified. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/2121 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/2121/1422