Expression of recombinant varicella-zoster virus gE Δ-Fc in CHO cells and analysis of its immunogenicity
10.3760/cma.j.cn112309-20200507-00245
- VernacularTitle:重组水痘-带状疱疹病毒gE Δ-Fc融合蛋白在CHO细胞中的表达、鉴定及免疫原性分析
- Author:
Wenyu CUI
1
;
Jilai LI
;
Zhuan ZHANG
;
Yibo HOU
;
Jing XU
Author Information
1. 国药中生生物技术研究院有限公司第三研究室，北京 101111
- From:
Chinese Journal of Microbiology and Immunology
2020;40(9):709-713
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express the recombinant varicella-zoster virus (VZV) gE Δ-Fc fusion protein using CHO cell expression system, and provide reference for screening candidate antigens of recombinant herpes zoster vaccines. Methods:A eukaryotic expression plasmid containing the gE Δ-Fc gene was transfected into CHO cells. Monoclonal cells were selected by methionine sulfoximine (MSX) pressure screening and limited dilution method to obtain the CHO cells secreting and expressing the gE Δ-Fc fusion protein. The expressed gE Δ-Fc fusion protein was purified by MabSelect SuRe affinity chromatography. The binding activity of gE Δ-Fc fusion protein to Fc receptors was identified by ELISA. Flow cytometry was used to detect the phagocytosis of antigens by DC2.4 cells. Antibody titers in serum samples of BALB/c mice immunized with the gE Δ-Fc fusion protein were detect by ELISA. Results:A CHO cell line stably expressing the gE Δ-Fc fusion protein was obtained. Flow cytometry suggested that the phagocytotic activity of DC2.4 cells against the gE Δ-Fc fusion protein was stronger than that against gE. Moreover, the gE Δ-Fc fusion protein could induce BALB/c mice to produce high titers of specific anti-VZV antibodies. Conclusions:The recombinant VZV gE Δ-Fc fusion protein expressed in CHO cells had a good immunogenicity. This study provided reference for screening candidate antigens of recombinant herpes zoster vaccines.
