Methods for enhancing the production of viral vaccines in cell culture
AssigneeThe Regents of the University of California
InventorAllan S. Lau
Life of this patent
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Abstract
Methods for enhancing the production of viral vaccines in animal cell culture are described. These methods rely on the manipulation of the cellular levels of certain interferon induced antiviral activities, in particular, cellular levels of double-stranded RNA (dsRNA) dependent kinase (PKR) and 2′-5′ oligoadenylate synthetase (2-5A synthetase). In cell cultures deficient for PKR or 2-5A synthetase, viral yield is enhanced by several orders of magnitude over cell cultures with normal levels of these proteins making these cell cultures useful for the production of viral vaccines.
Claims25
A method for production of a viral vaccine comprising:
- (a) selecting a cell used for the production of a vaccine;
- (b) making said cell deficient by targeted gene deletion in at least one interferon-stimulated gene (ISG) to render the cell deficient in the activity of the gene product of the interferon-stimulated gene, wherein the ISG is selected from the group consisting of a protein kinase RNA-dependent (PKR) gene, a 2′-5′-linked oligoadenylate (2-5A) synthetase gene, and an Mx gene, and wherein the deficient cell has decreased antiviral activity compared to the cell prior to modification;
- (c) infecting said deficient cell with a donor animal virus;
- (d) culturing said infected cell under conditions providing for efficient virus growth;
- (e) harvesting the virus produced;
- and
- (f) preparing said viral vaccine from said harvested virus.
A method for production of a viral vaccine comprising:
- (a) infecting a cell having a targeted deletion in a protein kinase RNA-dependent (PKR) gene with a donor animal virus, said targeted deletion rendering the cell deficient in PKR activity, wherein the cell has decreased antiviral activity compared to the cell prior to modification;
- (b) culturing said infected cell under conditions providing for efficient virus growth;
- (c) harvesting the virus produced;
- and
- (d) preparing said viral vaccine from said harvested virus.
Dependencies are inferred from the claim text.