Preparation, characterization and immunogenicity analysis of Chikungunya virus-like particles produced in insect cells
10.3760/cma.j.issn.1003-9279.2015.03.009
- VernacularTitle:基孔肯雅病毒样颗粒的制备及免疫原性研究
- Author:
Jiandong LI
1
;
Quanfu ZHANG
;
Shuo ZHANG
;
Chuan LI
;
Qinzhi LIU
;
Mifang LIANG
;
Dexin LI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Chikungunya virus;
Virus-like particles;
Baculoviridae;
Immunogenietices
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(3):219-221
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare the virus-like particles (VLPs) of Chikungunya virus (CHIKV) and evaluate the immunogenicity.Method CHIKV structural protein C-E3-E2-6K-E1 encoding gene were amplified by fusion PCR,and cloned into an insect cell expression vector.The recombinant Baculovirus were recovered by co-transfection of the expression plasmid with baculovirus linear DNA into SF9 insect cells,and CHIKV VLPs were prepared from suspension culture SF9 cells.Structural proteins expression were analyzed using IFA,SDS-PAGE and Western-Blot,and morphological analysis via electron microscopy.Result Which showed that CHIKV structural proteins were secreted into the cell culture supernatant and assembled into virus-like spherical particles.BALB /c mice were immunized with the VLPs,high levels of CHIKV specific antibodies were detected in the sera,and CHIKV infection of Vero cells could be effectively neutralized.Couclusion The results showed that CHIKV VLPs can be efficiently produced by the baculovirus expression system in insect cells,and specific IgG and neutralization antibodies could be induced in mice after VLPs immunization.This research laid the foundation for the development of CHIKV VLPs based on immunological detection reagents and even vaccines.
