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Process monitoring of virus-like particle reassembly by diafiltration with UV/Vis spectroscopy and light scattering

Rüdt, M.; Vormittag, P.; Hillebrandt, N.; Hubbuch, J.

Virus‐like particles (VLPs) have shown great potential as biopharmaceuticals in the marketand in clinics. Nonenveloped, in vivo assembled VLPs are typically disassembled andreassembled in vitro to improve particle stability, homogeneity, and immunogenicity. At theindustrial scale, cross‐flow filtration (CFF) is the method of choice for performingreassembly by diafiltration. Here, we developed an experimental CFF setup with an on‐linemeasurement loop for the implementation of process analytical technology (PAT). Themeasurement loop included an ultraviolet and visible (UV/Vis) spectrometer as well as alight scattering photometer. These sensors allowed for monitoring protein concentration,protein tertiary structure, and protein quaternary structure. The experimental setup wastested with three Hepatitis B core Antigen (HBcAg) variants. With each variant, threereassembly processes were performed at different transmembrane pressures (TMPs).While light scattering provided information on the assembly progress, UV/Vis allowed formonitoring the protein concentration and the rate of VLP assembly based on themicroenvironment of Tyrosine‐132. VLP formation was verified by off‐line dynamic lightscattering (DLS) and transmission electron microscopy (TEM). ... mehr

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DOI: 10.1002/bit.26935
Zitationen: 6
Web of Science
Zitationen: 6
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2019
Sprache Englisch
Identifikator ISSN: 0006-3592, 0368-1467, 1097-0290
KITopen-ID: 1000095179
Erschienen in Biotechnology & bioengineering
Band 116
Heft 6
Seiten 1366-1379
Vorab online veröffentlicht am 26.01.2019
Schlagwörter cross‐flow filtration, downstream processing, process analytical technology, self‐assembly, spectroscopy, virus‐like particles
Nachgewiesen in Scopus
Web of Science
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