Eukaryotic Expression and Immunogenic Research of Recombination Ebola Virus Membrane Protein Gp-Fc.
- Author:
Xiaoguang ZHANG
;
Ren YANG
;
Jiao WANG
;
Xuan WANG
;
Mieling HOU
;
Lina AN
;
Ying ZHU
;
Yuxi CAO
;
Yi ZENG
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
immunology;
Ebolavirus;
genetics;
immunology;
Female;
Gene Expression;
Hemorrhagic Fever, Ebola;
immunology;
virology;
Humans;
Immunization;
Male;
Mice;
Mice, Inbred BALB C;
Recombinant Proteins;
genetics;
immunology;
Viral Envelope Proteins;
genetics;
immunology
- From:
Chinese Journal of Virology
2016;32(1):8-13
- CountryChina
- Language:Chinese
-
Abstract:
We used 293 cells to express the recombinant membrane protein of the Ebola virus. Then, the immunogenicity of the recombinant protein was studied by immunized BALB/c mice. According to the codon use frequency of humans, the gene encoding the extracellular domain of the Ebola virus membrane protein was optimized, synthesized, and inserted into the eukaryotic expression plasmid pXG-Fc to construct the human IgG Fc and Ebola GP fusion protein expression plasmid pXG-modGP-Fc. To achieve expression, the fusion protein expression vector was transfected into high-density 293 cells using transient transfection technology. The recombinant protein was purified by protein A affinity chromatography. BALB/c mice were immunized with the purified fusion protein, and serum antibody titers evaluated by an indirect enzyme-linked immunosorbent assay (ELISA). Purification and analyses of the protein revealed that the eukaryotic expression vector could express the recombinant protein GP-Fc effectively, and that the recombinant protein in the supernatant of the cell culture was present as a dimer. After immunization with the purified recombinant protein, a high titer of antigen-specific IgG could be detected in the serum of immunized mice by indirect ELISA, showing that the recombinant protein had good immunogenicity. These data suggest that we obtained a recombinant protein with good immunogenicity. Our study is the basis for development of a vaccine against the Ebola virus and for screening of monoclonal antibodies.
