Expressing of Recombinant Vegfr2-Specific Nanobody in Baculoviruexpression System Publisher



Shokrollahi N¹ ; Habibianbouhi M² ; Jahaniannajafabadi A³ ; Alirahimi E¹ ; Behdani M¹ Show Affiliations
Source: Iranian Journal of Biotechnology Published:2021

Abstract

Background: Baculovirus expression system, introduced more than 20 years ago, is considered as a useful tool for large and complex eukaryotic recombinant protein production. A baculovirus expression vector is a recombinant virus which desired foreign protein coding sequences is under control of the virus gene promoter. Baculovirus only infects insect cells and do not normally infect vertebrates therefore, they possess no risk of biological risks for human. Objectives: The aim of this study was to recombinant expression of vascular endothelial growth factor (VEGF) reseptor-2 specific Nanobody in the baculovirus expression system. Materials and Methods: Gene of specific Nanobody against the VEGF reseptor-2 that called 3VGR19 was cloned and expressed in baculovirus system. Results: 3VGR19 Nanobody gene was amplified by Polymerase Chain Reaction (PCR) using the specific primers, and was cloned in pFastBac HTA plasmid. DH10Bac bacteria was transformed with resulted donor plasmid. The cultured Sf9 insect cell line was transfected with recombinant bacmid, and finally, the expression and purification of 3VGR19 was confirmed in insect cells. Conclusions: In conclusion, Transient infection of insect cells with baculovirus can be a promising technology for expression of antibody fragments. © 2021 The Author(s); Published by National Institute of Genetic Engineering and Biotechnology.