Improved aav production using potassium chloride
AssigneeUniversity of Florida Research Foundation, Inc.
InventorsNathalie Clement, Laura Adamson
Life of this patent
Estimating the term…
Abstract
Provided herein are methods of improving rAAV production in mammalian cells via the manipulation of a concentration of potassium chloride present in the media in which the mammalian cells are cultured and/or in which the rAAV particles are produced. The concentration of potassium chloride present in such media may be manipulated by, for example, supplementing a base medium with additional potassium chloride either before, at the same time as, or after the cells are contacted (e.g., infected) with one or more viral vectors (e.g., recombinant Herpes Simplex Virus (rHSV) vectors). Some embodiments of the present invention further contemplate supplementing the base medium with additional potassium chloride and sodium chloride in combination.
Claims81
A method of producing rAAV in mammalian cells, the method comprising:
- (i) culturing mammalian cells in a medium;
- (ii) adding one or more viral vectors encoding recombinant AAV (rAAV) genes and/or genes of interest into the medium;
- and
- (iii) supplementing the medium with at least 5 mM potassium chloride.
A method of producing rAAV in mammalian cells, the method comprising contacting mammalian cells with one or more viral vectors encoding recombinant AAV (rAAV) genes and/or genes of interest in the presence of at least 6 mM potassium chloride.
A method of producing rAAV in mammalian cells, the method comprising:
- (i) supplementing a medium with at least 5 mM potassium chloride;
- and
- (ii) contacting the mammalian cells with one or more viral vectors encoding recombinant AAV (rAAV) genes and/or genes of interest in the supplemented medium.
Dependencies are inferred from the claim text.