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Purification and assembling a fused capsid protein as an enterovirus 71 vaccine candidate from inclusion bodies to pentamer-based nanoparticles
Xue, Ling1; Liu, Jiangning2; Wang, Qi1,3; Zhang, Chun1; Xu, Longfu1,3; Luo, Jian1; Wang, Jian4; Qin, Chuan2; Liu, Yongdong1; Su, Zhiguo1
2017-01-15
Source PublicationBIOCHEMICAL ENGINEERING JOURNAL
ISSN1369-703X
Volume117Pages:139-146
Abstract

An efficient preparation process for a novel Enterovirus 71 (EV71) vaccine was developed in this paper, which is a fused antigen by connecting the truncated capsid proteins of VP1, VP2 and VP3 into one molecule through flexible peptide linkers and expressed in E. coli as inclusion bodies. The fused protein was purified at denatured state through two-step ion exchange chromatography, with final purity above 95%, host cell proteins below 0.003% and residual DNA less than 50 ng/mL. During the following refolding and assembling process through dilution, the fused antigen precipitated completely, while the precipitation was efficiently inhibited with 2 M urea or 0.5 M arginine as an additive. Size exclusion chromatography analysis indicated the protein formed soluble aggregates with linear or rod-like appearance in transmission electron microscopy. These aggregates transformed into pentamers with a size of 15 nm at pH 8.0 after the additive removing. Moreover, most of the pentamers assembled as sphere-like particulates about 25-40 nm after being induced by calcium chloride. High antigen-specific IgG titer was elicited by immunization with the nanoparticles in mouse model. Splenocytes proliferative responses and cytokines analysis indicated this particulate antigen could induce humoral and cellular immune responses. These results lay foundations for developing the fused antigen as an alternative vaccine against hand-foot-and-mouth disease (HFMD) and for the large-scale production for E.coli-based vaccines. (C) 2016 Elsevier B.V. All rights reserved.

KeywordEnterovirus 71 (Ev71) Virus-like Particles (Vlps) Assembling Fused Antigen Inclusion Bodies
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.bej.2016.10.009
Indexed BySCI
Language英语
WOS KeywordVirus-like Particles ; Mice ; Infection ; Vp1 ; Lethal ; Polyomavirus ; Immunization ; Expression ; Epitopes ; Dna
WOS Research AreaBiotechnology & Applied Microbiology ; Engineering
WOS SubjectBiotechnology & Applied Microbiology ; Engineering, Chemical
Funding OrganizationNational Science and Technology Major Project of
WOS IDWOS:000390629200017
Citation statistics
Cited Times:4[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/21861
Collection生化工程国家重点实验室
Affiliation1.Chinese Acad Sci, Natl Key Lab Biochem Engn, Inst Proc Engn, Beijing 100190, Peoples R China
2.Chinese Acad Med Sci, Inst Lab Anim Sci, Beijing 100021, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Natl Vaccine & Serum Inst Co Ltd, Beijing 100024, Peoples R China
Recommended Citation
GB/T 7714
Xue, Ling,Liu, Jiangning,Wang, Qi,et al. Purification and assembling a fused capsid protein as an enterovirus 71 vaccine candidate from inclusion bodies to pentamer-based nanoparticles[J]. BIOCHEMICAL ENGINEERING JOURNAL,2017,117:139-146.
APA Xue, Ling.,Liu, Jiangning.,Wang, Qi.,Zhang, Chun.,Xu, Longfu.,...&Su, Zhiguo.(2017).Purification and assembling a fused capsid protein as an enterovirus 71 vaccine candidate from inclusion bodies to pentamer-based nanoparticles.BIOCHEMICAL ENGINEERING JOURNAL,117,139-146.
MLA Xue, Ling,et al."Purification and assembling a fused capsid protein as an enterovirus 71 vaccine candidate from inclusion bodies to pentamer-based nanoparticles".BIOCHEMICAL ENGINEERING JOURNAL 117(2017):139-146.
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