US 12,146,149 B2
Method for adenovirus purification
Asa Hagner McWhirter, Uppsala (SE); and Anna Akerblom, Uppsala (SE)
Assigned to Cytiva BioProcess R&D AB, Uppsala (SE)
Appl. No. 17/047,843
Filed by Cytiva BioProcess R&D AB, Uppsala (SE)
PCT Filed Apr. 24, 2019, PCT No. PCT/EP2019/060431
§ 371(c)(1), (2) Date Oct. 15, 2020,
PCT Pub. No. WO2019/206940, PCT Pub. Date Oct. 31, 2019.
Claims priority of application No. 1806736 (GB), filed on Apr. 25, 2018.
Prior Publication US 2021/0292788 A1, Sep. 23, 2021
Int. Cl. C12N 15/86 (2006.01); C12N 7/00 (2006.01); C12N 15/861 (2006.01)
CPC C12N 15/86 (2013.01) [C12N 7/00 (2013.01); C12N 15/861 (2013.01); C12N 2710/10043 (2013.01); C12N 2710/10051 (2013.01)] 7 Claims
 
1. A method for adenovirus purification comprising the following steps: a) capturing adenovirus from an adenovirus-containing cell culture harvest on an anion exchanger resin, wherein the anion exchange resin comprises an agarose base matrix modified with a quaternary ammonium strong ion exchange ligand; b) eluting said adenovirus with a shallow conductivity gradient with an increasing salt concentration (mM) of 15-25% over the gradient; c) adding said eluted adenovirus to a shell bead resin comprising a porous, inert shell and a porous core, wherein the core is provided with hydrophobic interaction ligands; and d) eluting said adenovirus from said shell bead resin in the flow through, wherein the adenovirus eluted in step d) comprises less than 1 ng/ml host cell protein (HCP).