isolation of a novel anti-kdr3 singlechain variable fragment antibody from a phage display library

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.