Recombinant Expression of Hemagglutinin Protein of Iranian Swine Influenza a (H1n1) in the Insect Cells Using Baculovirus System Publisher



Zahmati S^{1, 4} ; Mahdavi M² ; Taghizadehtarnabi M³ ; Haghighat S^{1, 4} ; Jalalirad R⁵ Show Affiliations
Source: Journal of Isfahan Medical School Published:2021

Abstract

Background: Influenza virus, which is a decisive agent of influenza, attracted interest because of the event of the annual epidemic among a wide range of animal and human hosts. This study aimed to express the hemagglutinin protein of Iranian swine Influenza A (H1N1) through a baculovirus system in SF9 cells to produce a new recombinant vaccine. Methods: Hemagglutinin gene of Swine H1N1 virus was amplified with specific primers containing restriction enzymes site, and then cloned. Afterward, the vector was transformed to DH10Bac using Bac-to-Bac system in order to produce a recombinant bacmid. Hemagglutinin expression and its biological activity were assessed using molecular and immunization tests. Findings: The target rHA in length, 1710 bp, was produced and expressed in transfected SF9 cells with a size of ~66 kDa. The infected cells expanded in size and their nucleus, and desiccated from the surface of the cell culture as a granular. They could absorb chick red blood cells (RBCs), and appear as cell aggregates forty-eight post-infection. The fact that infected cells were unable to form cell clamp showed the test's accuracy and inhibition of hemodesorption activity. The amount of protein obtained was 10.76 µg/100 µl, equal to 0.1 mg/ml. Conclusion: The baculovirus expression system could express the recombinant protein in the insect cell. Therefore, it may be a well-suited alternative to produce a new generation of the vaccine instead of egg-based and cell-culture-based generation vaccines. © 2021 Isfahan University of Medical Sciences(IUMS). All rights reserved.