Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography

被引：21
|
作者：
Huhti, L. [1 ]
Tamminen, K. [1 ]
Vesikari, T. [1 ]
Blazevic, V. [1 ]
机构：
[1] Univ Tampere, Sch Med, Vaccine Res Ctr, Tampere 33520, Finland
来源：
ARCHIVES OF VIROLOGY | 2013年 / 158卷 / 05期
关键词：
ROTAVIRUS-LIKE PARTICLES; LARGE-SCALE PRODUCTION; NORWALK VIRUS; IMMUNE-RESPONSES; SPORADIC GASTROENTERITIS; SACCHAROMYCES-CEREVISIAE; MEMBRANE CHROMATOGRAPHY; VIRAL GASTROENTERITIS; INSECT CELLS; BACULOVIRUS;
D O I：
10.1007/s00705-012-1565-7
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
Recombinant baculovirus (BV) expression systems are widely applied in the production of viral capsid proteins and virus-like particles (VLPs) for use as immunogens and vaccine candidates. Traditional density gradient purification of VLPs does not enable complete elimination of BV-derived impurities, including live viruses, envelope glycoprotein gp64 and baculoviral DNA. We used an additional purification system based on ionic strength to purify norovirus (NoV) GII-4 capsid-derived VLPs. The anion exchange chromatography purification led to highly purified VLPs free from BV impurities with intact morphology. In addition, highly purified VLPs induced strong NoV-specific antibody responses in BALB/c mice. Here, we describe a method for NoV VLP purification and several methods for determining their purity, including quantitative PCR for BV DNA detection.
引用
收藏
页码：933 / 942
页数：10
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