Assessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing.

2.50
Hdl Handle:
http://hdl.handle.net/10033/566253
Title:
Assessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing.
Authors:
Zahid, Maria; Lünsdorf, Heinrich; Rinas, Ursula
Abstract:
The hepatitis B surface antigen (HBsAg) is a recombinant protein-based vaccine being able to form virus-like particles (VLPs). HBsAg is mainly produced using yeast-based expression systems, however, recent results strongly suggest that VLPs are not formed within the yeast cells during the cultivation but are formed in a gradual manner during the following down-stream procedures. VLPs are also not detectable during the first down-stream steps including mechanical and EDTA/detergent-assisted cell destruction. Moreover, VLPs are not detectable in the cell lysate treated with polyethylene glycol and colloidal silica. The first VLP resembling structures appear after elution of HBsAg from colloidal silica to which it binds through hydrophobic interaction. These first VLP resembling structures are non-symmetrical as well as heterodisperse and exhibit a high tendency toward cluster formation presumably because of surface exposed hydrophobic patches. More symmetrical and monodisperse VLPs appear after the following ion-exchange and size-exclusion chromatography most likely as the result of buffer changes during these purification steps (toward more neutral pH and less salt). Final treatment of the VLPs with the denaturant KSCN at moderate concentrations with following KSCN removal by dialysis does not cause unfolding and VLP disassembly but results in a re- and fine-structuring of the VLP surface topology.
Affiliation:
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Assessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing. 2015, 33 (31):3739-45 Vaccine
Journal:
Vaccine
Issue Date:
17-Jul-2015
URI:
http://hdl.handle.net/10033/566253
DOI:
10.1016/j.vaccine.2015.05.066
PubMed ID:
26079614
Type:
Article
Language:
en
ISSN:
1873-2518
Appears in Collections:
Publications of the RG Rekombinante Proteinexpression (RPEX)

Full metadata record

DC FieldValue Language
dc.contributor.authorZahid, Mariaen
dc.contributor.authorLünsdorf, Heinrichen
dc.contributor.authorRinas, Ursulaen
dc.date.accessioned2015-08-13T10:57:33Zen
dc.date.available2015-08-13T10:57:33Zen
dc.date.issued2015-07-17en
dc.identifier.citationAssessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing. 2015, 33 (31):3739-45 Vaccineen
dc.identifier.issn1873-2518en
dc.identifier.pmid26079614en
dc.identifier.doi10.1016/j.vaccine.2015.05.066en
dc.identifier.urihttp://hdl.handle.net/10033/566253en
dc.description.abstractThe hepatitis B surface antigen (HBsAg) is a recombinant protein-based vaccine being able to form virus-like particles (VLPs). HBsAg is mainly produced using yeast-based expression systems, however, recent results strongly suggest that VLPs are not formed within the yeast cells during the cultivation but are formed in a gradual manner during the following down-stream procedures. VLPs are also not detectable during the first down-stream steps including mechanical and EDTA/detergent-assisted cell destruction. Moreover, VLPs are not detectable in the cell lysate treated with polyethylene glycol and colloidal silica. The first VLP resembling structures appear after elution of HBsAg from colloidal silica to which it binds through hydrophobic interaction. These first VLP resembling structures are non-symmetrical as well as heterodisperse and exhibit a high tendency toward cluster formation presumably because of surface exposed hydrophobic patches. More symmetrical and monodisperse VLPs appear after the following ion-exchange and size-exclusion chromatography most likely as the result of buffer changes during these purification steps (toward more neutral pH and less salt). Final treatment of the VLPs with the denaturant KSCN at moderate concentrations with following KSCN removal by dialysis does not cause unfolding and VLP disassembly but results in a re- and fine-structuring of the VLP surface topology.en
dc.language.isoenen
dc.titleAssessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalVaccineen

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