Expression, purification, and characterization of a diabody against the most important angiogenesis cell receptor: Vascular endothelial growth factor receptor 2

Behdani, Mahdi; Zeinali, Sirous; Karimipour, Morteza; Khanahmad, Hossein; Asadzadeh, Nader; Azadmanesh, Kayhan; Seyed, Negar; Baniahmad, Seyed Farzad; Habibi Anbouhi, Mahdi

Expression, purification, and characterization of a diabody against the most important angiogenesis cell receptor: Vascular endothelial growth factor receptor 2

Authors

¹ Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran

² Department of Molecular Medicine, Pasteur Institute of Iran; Department of Anatomical Sciences, Division of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Iran

³ Department of Animal Breeding and Genetics, Animal Science Research Institute of Iran, Tehran, Iran

⁴ Department of Virology, Pasteur Institute of Iran, Tehran, Iran

Abstract

Antibodies and their derivative fragments have long been used as tools in a variety of applications, in fundamental research work, biotechnology, diagnosis, and therapy. Camels produce single heavy-chain antibodies (VHH) in addition to usual antibodies. These minimal-sized binders are very robust and bind the antigen with high affinity in a monomeric state. Vascular endothelial growth factor recepror-2 (VEGFR2) is an important tumor-associated receptor that blockade of its signaling can lead to the inhibition of neovascularization and tumor metastasis. Here, we describe the construction, expression, and purification VEGFR2-specific Diabody. Two variable fragments of a same camel anti-VEGFR2 antibody were linked together by the upper hinge segment of antibody to make a diabody. We showed the ability of diabody to recognition of VEGFR2 on the cell surface by FACS. Diabodies can be produced in the low-cost prokaryotic expression system, so they are suitable molecules for diagnostic and therapeutic issues.

Keywords

References

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