Isolation of a Novel Anti-Kdr3 Single-Chain Variable Fragment Antibody From a Phage Display Library Publisher Pubmed



Kordi S^{1, 2} ; Rahmatiyamchi M² ; Vostakolaei MA³ ; Etemadie A⁴ ; Barzegari A⁵ ; Abdolalizadeh J^{6, 7}
Source: Iranian Journal of Allergy# Asthma and Immunology Published:2019

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. Copyright © June 2019, Iran J Allergy Asthma Immunol. All rights reserved.