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Region Hovedstaden - en del af Københavns Universitetshospital
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Impact of the isoelectric point of model parvoviruses on viral retention in anion exchange chromatography

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  • Remo Leisi
  • Raphael Wolfisberg
  • Thomas Nowak
  • Oliver Caliaro
  • Andreas Hemmerle
  • Nathan J Roth
  • Carlos Ros
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Anion-exchange chromatography (AEX) is used in the downstream purification of monoclonal antibodies to remove impurities and potential viral contamination based on electrostatic interactions. Although the isoelectric point (pI) of viruses is considered a key factor predicting the virus adsorption to the resin, the precise molecular mechanisms involved remain unclear. To address this question, we compared structurally homologous parvoviruses that only differ in their surface charge distribution. A single charged amino acid substitution on the capsid surface of minute virus of mice (MVM) provoked an increased apparent pI (pI app) 6.2 compared to wild-type MVM (pI app = 4.5), as determined by chromatofocusing. Despite their radically different pI app, both viruses displayed the same interaction profile in Mono Q AEX at different pH conditions. In contrast, the closely related canine parvovirus (pI app = 5.3) displayed a significantly different interaction at pH 5. The detailed structural analysis of the intricate three-dimensional structure of the capsids suggests that the charge distribution is critical, and more relevant than the pI, in controlling the interaction of a virus with the chromatographic resin. This study contributes to a better understanding of the molecular mechanisms governing virus clearance by AEX, which is crucial to enable robust process design and maximize safety.

OriginalsprogEngelsk
TidsskriftBiotechnology and bioengineering
ISSN0006-3592
DOI
StatusUdgivet - 4 sep. 2020

ID: 60791558