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

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,...

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Detalles Bibliográficos
Autores principales: Huhti, L., Tamminen, K., Vesikari, T., Blazevic, V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Vienna 2012
Materias:
Original Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087179/
https://www.ncbi.nlm.nih.gov/pubmed/23229011
http://dx.doi.org/10.1007/s00705-012-1565-7
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author Huhti, L.
Tamminen, K.
Vesikari, T.
Blazevic, V.
author_facet Huhti, L.
Tamminen, K.
Vesikari, T.
Blazevic, V.
author_sort Huhti, L.
collection PubMed
description 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.
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spelling pubmed-70871792020-03-23 Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography Huhti, L. Tamminen, K. Vesikari, T. Blazevic, V. Arch Virol Original Article 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. Springer Vienna 2012-12-11 2013 /pmc/articles/PMC7087179/ /pubmed/23229011 http://dx.doi.org/10.1007/s00705-012-1565-7 Text en © Springer-Verlag Wien 2012 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Article
Huhti, L.
Tamminen, K.
Vesikari, T.
Blazevic, V.
Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title_full Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title_fullStr Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title_full_unstemmed Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title_short Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
title_sort characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087179/
https://www.ncbi.nlm.nih.gov/pubmed/23229011
http://dx.doi.org/10.1007/s00705-012-1565-7
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