Details
| Original language | English |
|---|---|
| Pages (from-to) | 3739-3745 |
| Number of pages | 7 |
| Journal | Vaccine |
| Volume | 33 |
| Issue number | 31 |
| Publication status | Published - 17 Jul 2015 |
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.
Keywords
- Aerosil-380, Hepatitis B surface antigen virus-like particles, Ion-exchange chromatography, Purification, Size-exclusion chromatography, Transmission electron microscopy, Vaccine
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Immunology and Microbiology(all)
- veterinary(all)
- Medicine(all)
- Public Health, Environmental and Occupational Health
- Medicine(all)
- Infectious Diseases
Sustainable Development Goals
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In: Vaccine, Vol. 33, No. 31, 17.07.2015, p. 3739-3745.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Assessing stability and assembly of the hepatitis B surface antigen into virus-like particles during down-stream processing
AU - Zahid, Maria
AU - Lünsdorf, Heinrich
AU - Rinas, Ursula
N1 - Funding Information: Maria Zahid wishes to express her gratitude to the Deutscher Akademischer Austauschdienst (DAAD) of Germany and the Higher Education Commission (HEC) of Pakistan for her fellowship. Publisher Copyright: © 2015 Elsevier Ltd.
PY - 2015/7/17
Y1 - 2015/7/17
N2 - 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.
AB - 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.
KW - Aerosil-380
KW - Hepatitis B surface antigen virus-like particles
KW - Ion-exchange chromatography
KW - Purification
KW - Size-exclusion chromatography
KW - Transmission electron microscopy
KW - Vaccine
UR - http://www.scopus.com/inward/record.url?scp=84937635544&partnerID=8YFLogxK
U2 - 10.1016/j.vaccine.2015.05.066
DO - 10.1016/j.vaccine.2015.05.066
M3 - Article
C2 - 26079614
AN - SCOPUS:84937635544
VL - 33
SP - 3739
EP - 3745
JO - Vaccine
JF - Vaccine
SN - 0264-410X
IS - 31
ER -