Expression and purification of the chimeric virus-like particles displaying enterovirus 71 epitopes
10.3760/cma.j.issn.1003-9279.2018.02.020
- VernacularTitle: EV71中和表位嵌合病毒样颗粒的表达纯化
- Author:
Pu LIANG
1
;
Yao YI
;
Qiudong SU
;
Shengli BI
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
- Publication Type:Journal Article
- Keywords:
Enterovirus 71;
Virus-like particles;
Hepatitis B virus core antigen;
SP70;
Protein purification
- From:
Chinese Journal of Experimental and Clinical Virology
2018;32(2):199-202
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the expression and purification of the chimeric virus-like particles displaying epitopes of EV71 as a candidate of enterovirus 71 gene recombined vaccinet.
Methods:The fusion protein hepatitis B core (HBc)-SP70 was constructed by inserting SP70 into the main immunogenic region of truncated hepatitis B core antigen (HBcAg) sequence, expressed in E. Coli, and purified through sonication, ion exchange chromatography, CsCl cushion centrifugation and density gradient centrifugation. Then its antigenicity was detected by ELISA and Western blot assay.
Results:Recombinant plasmid pHBc-SP70 was successfully constructed. And the soluble fusion protein was efficiently expressed induced by IPTG. The purity of the chimeric virus-like particles (VLPs) was up to 90% after the purification process described in method . The purified fusion protein HBc-SP70 could be spontaneously folded and assembled into empty virus-like particles and react with the monoclonal antibodies against HBc and SP70.
Conclusions:The chimeric VLPs displaying epitopes of EV71 were efficiently expressed and purified in E. Coli. with excellent antigenicity, which laid a foundation for evaluation of the immune effect evoked by this EV71 gene recombined vaccine.
