Isolation of a Novel Anti-KDR3 Single-chain Variable Fragment Antibody from a Phage Display Library

  • 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 http://orcid.org/0000-0002-8283-5057
Keywords: Monoclonal antibody, Kinase insert domain receptor 3 (KDR3), Phage display, Single-chain variable fragment (SCFV), Vascular endothelial growth factor receptor-2 (VEGFR2)

Abstract

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.

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Published
2019-06-08
How to Cite
1.
Kordi S, Rahmati-Yamchi M, Asghari Vostakolaei M, Etemadie A, Barzegari A, Abdolalizadeh J. Isolation of a Novel Anti-KDR3 Single-chain Variable Fragment Antibody from a Phage Display Library. Iran J Allergy Asthma Immunol. 18(3):289-299.
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Original Article(s)