Purification of vaccinia viruses using hydrophobic interaction chromatography
AssigneesBavarian Nordic A/S, Otto-von-Guericke-Universität, Sartorius Stedim Biotech GmbH
InventorsSara Post Hansen, Rene Faber, Udo Reichl, Michael Wolff, Anders Peter Gram
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Abstract
The present invention relates to methods for purification of Vaccinia viruses (VV) and/or Vaccinia virus (VV) particles, which can lead to highly pure and stable virus preparations of predominantly biologically active viruses. The invention encompasses purifying a virus preparation in a sterilized way with high efficiency and desirable yield in terms of purity, biological activity and stability, aspects advantageous for industrial production.
Claims22
An industrial-scale method for the purification of biologically active Vaccinia viruses comprising:
- i) binding the Vaccinia viruses to a solid-phase sulfated cellulose matrix;
- ii) eluting the Vaccinia viruses;
- iii) binding the eluted Vaccinia viruses to a solid-phase hydrophobic interaction chromatograpy (HIC) matrix comprising a phenyl ligand;
- and
- iv) eluting a minimum of 5.0×1012 Vaccinia virus particles.
An industrial-scale method for the purification of biologically active Vaccinia viruses comprising:
- i) binding the Vaccinia viruses to a solid-phase heparin ligand matrix;
- ii) eluting the Vaccinia viruses;
- iii) binding the eluted Vaccinia viruses to a solid-phase hydrophobic interaction chromatograpy (HIC) matrix comprising a phenyl ligand;
- and
- iv) eluting a minimum of 5.0×1012 Vaccinia virus particles.
Dependencies are inferred from the claim text.