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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
发表期刊BIOCHEMICAL ENGINEERING JOURNAL
ISSN1369-703X
卷号117页码:139-146
摘要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.
关键词Enterovirus 71 (Ev71) Virus-like Particles (Vlps) Assembling Fused Antigen Inclusion Bodies
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.bej.2016.10.009
收录类别SCI
语种英语
关键词[WOS]VIRUS-LIKE PARTICLES ; MICE ; INFECTION ; VP1 ; LETHAL ; POLYOMAVIRUS ; IMMUNIZATION ; EXPRESSION ; EPITOPES ; DNA
WOS研究方向Biotechnology & Applied Microbiology ; Engineering
WOS类目Biotechnology & Applied Microbiology ; Engineering, Chemical
项目资助者National Science and Technology Major Project of "National Key Program on Drug Innovation"(2012ZX09101319004 ; National Natural Science Foundation of China(21576267) ; NN-CAS research fund(NN-CAS-2014-2) ; Open Funding Project of the National Key Laboratory of Biochemical Engineering(2014KF-05) ; 2016ZX09101120-006)
WOS记录号WOS:000390629200017
引用统计
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/21861
专题生化工程国家重点实验室
作者单位1.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
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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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