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Process development for cross-flow diafiltration-based VLP disassembly: A novel high-throughput screening approach

Hillebrandt, Nils 1; Vormittag, Philipp 1; Dietrich, Annabelle 1; Wegner, Christina H. 1; Hubbuch, Jürgen ORCID iD icon 1
1 Karlsruher Institut für Technologie (KIT)

Abstract:

Virus-like particles (VLPs) are particulate structures, which are applied as vaccines or delivery vehicles. VLPs assemble from subunits, named capsomeres, composed of recombinantly expressed viral structural proteins. During downstream processing, in vivo-assembled VLPs are typically dis- and reassembled to remove encapsulated impurities and to improve particle morphology. Disassembly is achieved in a high-pH solution and by the addition of a denaturant or reducing agent. The optimal disassembly conditions depend on the VLP amino acid sequence and structure, thus requiring material-consuming disassembly experiments. To this end, we developed a low-volume and high-resolution disassembly screening that provides time-resolved insight into the VLP disassembly progress. In this study, two variants of C-terminally truncated hepatitis B core antigen were investigated showing different disassembly behaviors. For both VLPs, the best capsomere yield was achieved at moderately high urea concentration and pH. Nonetheless, their disassembly behaviors differed particularly with respect to disassembly rate and aggregation. Based on the high-throughput screening results, a diafiltration-based disassembly process step was developed. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000136002
Veröffentlicht am 30.07.2021
Originalveröffentlichung
DOI: 10.1002/bit.27868
Scopus
Zitationen: 5
Web of Science
Zitationen: 5
Dimensions
Zitationen: 6
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2021
Sprache Englisch
Identifikator ISSN: 0006-3592, 0368-1467, 1097-0290, 1547-173X
KITopen-ID: 1000136002
Erschienen in Biotechnology & bioengineering
Verlag John Wiley and Sons
Band 118
Heft 10
Seiten 3926-3940
Vorab online veröffentlicht am 25.06.2021
Nachgewiesen in Scopus
Web of Science
Dimensions
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