| Expression, purification and characterization of enterovirus-71 virus-like particles. | |
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MedLine Citation:
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PMID: 16521221 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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AIM: Enterovirus 71 (EV71) has been implicated as the etiological agent responsible for the recent outbreaks of hand, foot and mouth disease associated with severe neurological diseases in the Asia-Pacific region. METHODS: The assembly process was hypothesized to occur via an orchestrated proteolytic processing of the P1 precursor by the viral protease 3CD. To test this hypothesis, we constructed 3 recombinant baculoviruses: Bac-P1 expressing P1; Bac-3CD expressing 3CD; and Bac-P1-3CD co-expressing P1 and 3CD. RESULTS: Both single infection by Bac-P1-3CD and co-infection by Bac-P1 and Bac-3CD resulted in correct cleavage of P1 to yield individual proteins VP0, VP1 and VP3, while the former approach yielded higher VLP production. In the cells, the structural proteins self-assembled into clusters of virus-like particles (VLP) resembling the authentic EV71 particle aggregates. After ultracentrifugation purification, the dispersed VLPs were indistinguishable from the authentic virus in size, appearance, composition and surface epitopes, as determined by SDS-PAGE, Western blot, transmission electron microscopy and immunogold labeling. CONCLUSION: Our data, for the first time, suggest that in insect cells EV71 structural proteins adopt a processing and assembly pathway similar to poliovirus assembly. The preservation of particle morphology and composition suggest that the VLP may be a valuable vaccine candidate to prevent EV71 epidemics. |
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Authors:
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Yao-Chi Chung; Jen-Huang Huang; Chia-Wei Lai; Heng-Chun Sheng; Shin-Ru Shih; Mei-Shang Ho; Yu-Chen Hu |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: World journal of gastroenterology : WJG Volume: 12 ISSN: 1007-9327 ISO Abbreviation: World J. Gastroenterol. Publication Date: 2006 Feb |
Date Detail:
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Created Date: 2006-03-07 Completed Date: 2006-05-02 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 100883448 Medline TA: World J Gastroenterol Country: China |
Other Details:
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Languages: eng Pagination: 921-7 Citation Subset: IM |
Affiliation:
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Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan, China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Disease Outbreaks Enterovirus / classification*, isolation & purification*, physiology Enterovirus Infections / epidemiology Humans Spodoptera Viral Proteins / genetics |
| Chemical | |
Reg. No./Substance:
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0/Viral Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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