Elsevier

Journal of Biotechnology

Volume 323, 10 November 2020, Pages 62-72
Journal of Biotechnology

Selection of chromatographic methods for the purification of cell culture-derived Orf virus for its application as a vaccine or viral vector

https://doi.org/10.1016/j.jbiotec.2020.07.023Get rights and content

Highlights

  • Estimation of the isoelectric point and size of Vero cell-derived Orf virus.

  • Limited dynamic binding capacity of tested Orf virus to sulfated cellulose.

  • Purification of Orf virus by steric exclusion chromatography lead to 84 % recovery.

  • Hydrophobic interaction chromatography suitable for Orf virus purification.

  • Promising unit operations for a scalable DSP to produce Orf virus viral vectors.

Abstract

In recent years, the Orf virus has become a promising tool for protective recombinant vaccines and oncolytic therapy. However, suitable methods for an Orf virus production, including up- and downstream, are very limited. The presented study focuses on downstream processing, describing the evaluation of different chromatographic unit operations. In this context, ion exchange-, pseudo-affinity- and steric exclusion chromatography were employed for the purification of the cell culture-derived Orf virus, aiming at a maximum in virus recovery and contaminant depletion. The most promising chromatographic methods for capturing the virus particles were the steric exclusion- or salt-tolerant anion exchange membrane chromatography, recovering 84 % and 86 % of the infectious virus. Combining the steric exclusion chromatography with a subsequent Capto™ Core 700 resin or hydrophobic interaction membrane chromatography as a secondary chromatographic step, overall virus recoveries of up to 76 % were achieved. Furthermore, a complete cellular protein removal and a host cell DNA depletion of up to 82 % was possible for the steric exclusion membranes and the Capto™ Core 700 combination.

The study reveals a range of possible unit operations suited for the chromatographic purification of the cell culture-derived Orf virus, depending on the intended application, i.e. a human or veterinary use, and the required purity.

Abbreviations

DBC
dynamic binding capacity
DLS
dynamic light scattering
HIC
hydrophobic interaction chromatography
HICP
HIC membrane adsorber with phenyl ligand
IEX
ion exchange chromatography
IEX-Q
strong anion exchanger (Quaternary ammonium)
IEX-S
strong cation exchanger (Methyl sulfonate)
IEX-STPA
salt tolerant polyamide anion exchanger
IU
infective units
MVA
Modified Vaccinia Ankara virus
PEG
polyethylene glycol
pI
isoelectric point
SCMA
sulfated cellulose membrane adsorber
SEC
size exclusion chromatography
SXC
steric exclusion chromatography
TCID50
fifty-percent tissue culture infective dose

Keywords

Parapoxvirus
Viral vector
Steric exclusion chromatography
Isoelectric point

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