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Green, Nicola K
Bracewell, Daniel G
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AbstractViral vectors such as adenovirus have successful applications in vaccines and gene therapy but the manufacture of the high‐quality virus remains a challenge. It is desirable to use the adsorption‐based chromatographic separations that so effectively underpin the therapeutic protein manufacture. However fundamental differences in the size and stability of this class of product mean it is necessary to revisit the design of sorbent's morphology and surface chemistry. In this study, the behaviour of a cellulose nanofiber ion‐exchange sorbent derivatised with quaternary amine ligands at defined densities is characterised to address this. This material was selected as it has a large accessible surface area for viral particles and rapid process times. Initially, the impact of surface chemistry on infective product recovery using low (440 µmol/g), medium (750 µmol/g), and high (1029 µmol/g) ligand densities is studied. At higher densities product stability is reduced, this effect increased with prolonged adsorption durations of 24 min with just ~10% loss at low ligand density versus ~50% at high. This could be mitigated by using a high flow rate to reduce the cycle time to ~1 min. Next, the impact of ligand density on the separation's resolution was evaluated. Key to understanding virus quality is the virus particle: infectious virus particle ratio. It was found this parameter could be manipulated using ligand density and elution strategy. Together this provides a basis for viral vector separations that allows for their typically low titres and labile nature by using high liquid velocity to minimise both load and on‐column times while separating key product and process‐related impurities.
CitationTurnbull, J., Wright, B., Green, N. K., Tarrant, R., Roberts, I., Hardick, O. and Bracewell, D. G. (2019) Adenovirus 5 recovery using nanofiber ion‐exchange adsorbents, Biotechnology and Bioengineering, 116(7), pp. 1698-1709.
JournalBiotechnology and Bioengineering
DescriptionThis is an accepted manuscript of an article published by Wiley in Biotechnology and Bioengineering on 28/03/2019, available online: https://doi.org/10.1002/bit.26972 The accepted version of the publication may differ from the final published version.
SponsorsThis study was supported by the UK Engineering and Physical Sciences Research Council (EPSRC) grant EP/L01520X/1, and Puridify (now part of GE Healthcare), in conjunction with grant EP/N013395/1.
Except where otherwise noted, this item's license is described as https://creativecommons.org/licenses/by-nc-nd/4.0/