Method for purifying virus
AssigneeOxford University Innovation Limited
InventorsAlexander DOUGLAS, Carina Citra Dewi JOE
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Abstract
The invention relates to a method for purifying an adenovirus comprising (a) providing a liquid sample comprising adenovirus, (b) clarifying said sample by depth filtration, (c) performing anion exchange chromatography comprising the steps of (i) directly applying the clarified sample of (b) to an anion exchange column, (ii) eluting adenovirus from the anion exchange column to provide an eluate. The invention also relates to adenovirus produced from said methods, and to compositions comprising same.
Claims29
A method for purifying an adenovirus comprising
- (a) providing a liquid sample comprising adenovirus
- (b) clarifying said sample by depth filtration
- (c) performing anion exchange chromatography comprising the steps of
- (i) directly applying the clarified sample of (b) to anion exchange
- (ii) eluting adenovirus from the anion exchange to provide an eluate.
A method according to any preceding claim wherein the anion exchange chromatography is carried out with load and wash salt concentrations in the range 24 to 31 mS/cm, with 20 mM Tris pH 8.0.
A method according to any preceding claim wherein the anion exchange chromatography is carried out with load and wash salt concentrations in the range 280 mM NaCl to 361 mM NaCl, with 20 mM Tris pH 8.0.
A method according to any preceding claim wherein the anion exchange chromatography is carried out with wash conditions of 15 mS/cm to 30 mS/cm conductivity, with 20 mM Tris pH 8.0.
A method according to any preceding claim wherein the sample is adjusted to a conductivity of 28 mS/cm before applying to the anion exchange column.
A method according to any preceding claim wherein said depth filtration of step (b) comprises primary clarification followed by secondary clarification.
A method according to any preceding claim wherein said depth filtration of step (b) comprises combined primary and secondary clarification.
A method according to any preceding claim wherein said depth filtration of step (b) comprises use of a CoSP depth filter.
A method according to any preceding claim wherein the anion exchange chromatography of (c) is performed in-line with the clarifying depth filtration of (b) as a single unit operation.
A method according to any preceding claim wherein said liquid sample comprises cell lysate produced from cultured host cells comprising adenovirus.
A method according to any preceding claim further comprising:
- (d) performing buffer exchange on the eluate of (c).
A method according to any preceding claim wherein said adenovirus is, or is derived from, a simian adenovirus, preferably wherein said adenovirus is, or is derived from, a species E simian adenovirus, preferably wherein said adenovirus is ChAdOx1.
An adenovirus prepared by a method according to any preceding claim.
Dependencies are inferred from the claim text.