LAPSE:2020.0211
Published Article
LAPSE:2020.0211
A Flow-Through Chromatographic Strategy for Hepatitis C Virus-Like Particles Purification
Ricardo J. S. Silva, Mafalda G. Moleirinho, Ana S. Moreira, Alex Xenopoulos, Paula M. Alves, Manuel J. T. Carrondo, Cristina Peixoto
February 12, 2020
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).
Keywords
downstream processing, flow-through purification, hepatitis C, radial-flow chromatography, virus-like particles
Suggested Citation
Silva RJS, Moleirinho MG, Moreira AS, Xenopoulos A, Alves PM, Carrondo MJT, Peixoto C. A Flow-Through Chromatographic Strategy for Hepatitis C Virus-Like Particles Purification. (2020). LAPSE:2020.0211
Author Affiliations
Silva RJS: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal
Moleirinho MG: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
Moreira AS: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
Xenopoulos A: EMD Millipore Corporation, 80 Ashby Road, Bedford, MA 01730, USA
Alves PM: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
Carrondo MJT: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal
Peixoto C: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
Journal Name
Processes
Volume
8
Issue
1
Article Number
E85
Year
2020
Publication Date
2020-01-08
Published Version
ISSN
2227-9717
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Original Submission
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PII: pr8010085, Publication Type: Journal Article
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LAPSE:2020.0211
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doi:10.3390/pr8010085
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Feb 12, 2020
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Calvin Tsay
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