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Open AccessArticle

A Flow-Through Chromatographic Strategy for Hepatitis C Virus-Like Particles Purification

1
iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal
2
ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
3
EMD Millipore Corporation, 80 Ashby Road, Bedford, MA 01730, USA
*
Author to whom correspondence should be addressed.
Processes 2020, 8(1), 85; https://doi.org/10.3390/pr8010085
Received: 4 December 2019 / Revised: 27 December 2019 / Accepted: 3 January 2020 / Published: 8 January 2020
(This article belongs to the Section Biological Systems)
Biopharmaceuticals are currently becoming one of the fastest growing segments of the global pharmaceutical industry, being used in practically all branches of medicine from disease treatment to prevention. Virus-like particles (VLP) hold tremendous potential as a vaccine candidate due to their anticipated immunogenicity and safety profile when compared to inactivated or live attenuated viral vaccines. Nevertheless, there are several challenges yet to be solved in the development and manufacturing of these products, which ultimately can increase time to market. Suchlike virus-based products, the development of a platform approach is often hindered due to diversity and inherent variability of physicochemical properties of the product. In the present work, a flow-through chromatographic purification strategy for hepatitis C VLP expressed using the baculovirus-insect cell expression system was developed. The impact of operational parameters, such as residence time and ionic strength were studied using scaled-down models and their influence on the purification performance was described. The flow-through strategy herein reported made use of radial-flow chromatography columns packed with an anion exchanger and was compared with a bind and elute approach using the same chromatography media. Overall, by selecting the optimal operational setpoints, we were able to achieve higher VLP recoveries in the flow-through process (66% versus 37%) with higher removal of DNA, baculovirus and host-cell protein (92%, 99% and 50% respectively). View Full-Text
Keywords: downstream processing; flow-through purification; hepatitis C; radial-flow chromatography; virus-like particles downstream processing; flow-through purification; hepatitis C; radial-flow chromatography; virus-like particles
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MDPI and ACS Style

Silva, R.J.S.; Moleirinho, M.G.; Moreira, A.S.; Xenopoulos, A.; Alves, P.M.; Carrondo, M.J.T.; Peixoto, C. A Flow-Through Chromatographic Strategy for Hepatitis C Virus-Like Particles Purification. Processes 2020, 8, 85.

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