Display of B-cell linear epitopes of SARS-CoV-2 spike protein with hepatitis B virus core particles and analysis of their immunogenicity
10.3760/cma.j.cn112309-20230307-00052
- VernacularTitle:乙型肝炎病毒核心颗粒展示新型冠状病毒刺突蛋白B细胞线性表位及其免疫原性分析
- Author:
Xiaomei CHEN
1
;
Wenjie YUAN
;
Shijie WANG
;
Jianrong WANG
;
Xiaofeng HANG
;
Yanmin WAN
;
Yongxiang WANG
Author Information
1. 复旦大学上海医学院基础医学院病原生物学系，教育部、卫健委、医科院医学分子病毒学重点实验室，上海 200032
- Keywords:
Immunogenicity;
SARS-CoV-2;
B-cell linear epitope;
Capsid-like particles;
Antibody
- From:
Chinese Journal of Microbiology and Immunology
2024;44(3):234-240
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To obtain hepatitis B virus capsid-like particles (CLPs) displaying B-cell linear epitopes of SARS-CoV-2 spike protein and evaluate their immunogenicity.Methods:Four recombinant plasmids expressing fusion proteins (M1-HBc, S1-57-HBc, S14P5-HBc and S21P2-HBc) were constructed by separately replacing codon of alanine at position 80 of hepatitis B virus core protein (HBc) with four genes coding for four B-cell linear epitopes (M1, S1-57, S14P5 and S21P2). These four recombinant proteins were expressed in E. coli BL21 Star (DE3) strains. The expression products were identified using SDS-PAGE, Western blot and native agarose gel electrophoresis (NAGE). CLPs were purified by sucrose density gradient ultracentrifugation, verified for antigenicity by Western blot and used to immunize BALB/c mice. Serum antibody titers were detected by ELISA. Results:The recombinant fusion proteins M1-HBc and S1-57-HBc self-assembled into M1-CLP and S1-57-CLP. The titer of antibody against S1-57 polypeptide in S1-57-CLP-immunized mouse serum approached 1∶1 000 000.Conclusions:Hepatitis B virus CLPs displaying SARS-CoV-2 M1 or S1-57 linear epitopes are successfully expressed in a prokaryotic system and purified. S1-57-CLP has good immunogenicity. This study provides a new idea for the development of novel diagnostic reagents and vaccines for SARS-CoV-2.
