Shirafkan Kordi
1,2 
, Mohammad Rahmati-Yamchi
2 , Mehdi Asghari Vostakolaei
3,4, Abolfazl Barzegari
5 , Jalal Abdolalizadeh
6,7* 1 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
5 Research Centre for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran.
6 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
7 Paramedical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract
Purpose: The single-chain variable fragment (scFv) domain of antibodies is now considered asone of the therapeutic tools that can be produced by phage display technology (PDT). Antibodypurification is one of the most important steps in antibodies production. The aim of study waspurification and characterization of anti-VEGFR2 scFv antibody fragments.Methods: After the coating of vascular endothelial growth factor receptor 2 (VEGFR2) peptidein ELISA microplates, the phage display library of Tomlinson was used for antibody isolation.The targeted scFv was purified by chromatography using a zeolite-based column. The purity andfunctional assessment of purified scFv were evaluated by sodium dodecyl sulfate polyacrylamidegel electrophoresis (SDS-PAGE) and western blotting techniques, respectively. Affinity bindingwas evaluated by surface plasmon resonance (SPR).Results: The desired scFv was selected after four stages of biopanning. SDS-PAGE analysisshowed a 28 kDa scFv with high purity (>90%). The western bloting analysis confirmed thebinding of produced scFv antibody to the desired peptide. The affinity binding of scFv antibodyanalyzed by SPR was about 60 μM.Conclusion: In this study, the novel scFv antibody against VEGFR2 peptide was purified bychromatography column containing zeolite. Based on our findings the produced antibody maybe applied for diagnosis or targeting of VEGFR2 in antibody-based therapy strategies.