A flow-through chromatographic strategy for hepatitis c virus-like particles purification

Ricardo Jorge Sousa da Silva, Mafalda G. Moleirinho, Ana S. Moreira, Alex Xenopoulos, Paula M. Alves, Manuel J.T. Carrondo, Cristina Peixoto

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)


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).

Original languageEnglish
Article number85
Issue number1
Publication statusPublished - 1 Jan 2020


  • Downstream processing
  • Flow-through purification
  • Hepatitis C
  • Radial-flow chromatography
  • Virus-like particles


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