title: Characterization and immunogenicity of norovirus capsid-derived virus-like particles purified by anion exchange chromatography.
abstract: 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.
authors.affiliation: Vaccine Research Center, University of Tampere Medical School, Biokatu 10, 33520 Tampere, Finland. leena.huhti@uta.fi
authors.forename: L
authors.initials: L
authors.lastname: Huhti
authors.forename: K
authors.initials: K
authors.lastname: Tamminen
authors.forename: T
authors.initials: T
authors.lastname: Vesikari
authors.forename: V
authors.initials: V
authors.lastname: Blazevic
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology* / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D029322:Norovirus / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014765:Viral Vaccines