Expression, purification, characterization and immunogenicity of human immunodeficiency virus-1 glycoprotein gp120 derived from insect cells
10.3760/cma.j.issn.0254-5101.2017.09.001
- VernacularTitle:HIV-1糖蛋白gp120在昆虫细胞中的表达、纯化、性质鉴定以及免疫原性分析
- Author:
Zhenyong ZHANG
1
;
Tingting LI
;
Jiaming QIAO
;
Yuyun ZHANG
;
Shuangquan GAO
;
Qiaobin YAO
;
Zekai LI
;
Zhiqing ZHANG
;
Ying GU
;
Shaowei NINGSHAO
;
Li XIA
Author Information
1. 生命科学学院
- Keywords:
Human immunodeficiency virus-1;
Insect cell expression system;
Glycoprotein;
gp120;
Immune response
- From:
Chinese Journal of Microbiology and Immunology
2017;37(9):645-649
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.
