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Integrated Process for Capture and Purification of Virus-Like Particles: Enhancing Process Performance by Cross-Flow Filtration

Hillebrandt, N.; Vormittag, P.; Bluthardt, N.; Dietrich, A.; Hubbuch, J.

Abstract (englisch):
Virus-like particles (VLPs) are emerging nanoscale protein assemblies applied as prophylactic vaccines and in development as therapeutic vaccines or cargo delivery systems. Downstream processing (DSP) of VLPs comes both with challenges and opportunities, depending on the complexity and size of the structures. Filtration, precipitation/re-dissolution and size-exclusion chromatography (SEC) are potent technologies exploiting the size difference between product and impurities. In this study, we therefore investigated the integration of these technologies within a single unit operation, resulting in three different processes, one of which integrates all three technologies. VLPs, contained in clarified lysate from Escherichia coli, were precipitated by ammonium sulfate, washed, and re-dissolved in a commercial cross-flow filtration (CFF) unit. Processes were analyzed for yield, purity, as well as productivity and were found to be largely superior to a reference centrifugation process. Productivity was increased 2.6-fold by transfer of the wash and re-dissolution process to the CFF unit. Installation of a multimodal SEC column in the permeate line increased purity to 96% while maintaining a high productivity and high yield of 86%. ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/1000120987
Veröffentlicht am 07.07.2020
Originalveröffentlichung
DOI: 10.3389/fbioe.2020.00489
Scopus
Zitationen: 1
Web of Science
Zitationen: 1
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2020
Sprache Englisch
Identifikator ISSN: 2296-4185
KITopen-ID: 1000120987
Erschienen in Frontiers in Bioengineering and Biotechnology
Band 8
Seiten Aericle No.489
Vorab online veröffentlicht am 25.05.2020
Nachgewiesen in Web of Science
Scopus
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