Scalable method for recombinant AAV production
AssigneeREGENXBIO Inc.
InventorsMichael Gillmeister, Miguel Valle, Robert Stadelman, Franz Gerner, Matthieu Guibert, Bhargavi Kondragunta
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Abstract
Provided herein are improved methods for producing recombinant Adeno-Associated Virus (rAAV) particles. In some embodiments, a method for producing recombinant AAV (rAAV) particles provided herein comprises culturing cells capable of producing rAAV particles in the presence of a histone deacetylase (HDAC) inhibitor. In some embodiments, a method for producing recombinant AAV (rAAV) particles provided herein comprises culturing cells capable of producing rAAV particles in the presence of a histone deacetylase (HDAC) inhibitor and increased amount of a sodium salt.
Claims14
A method for producing recombinant adeno associated virus (rAAV) particles, comprising
- a) providing a cell culture comprising a cell capable of producing rAAV and transfecting the cell with one or more polynucleotides comprising or encoding
- an rAAV genome to be packaged,
- adenovirus helper functions necessary for packaging,
- an AAV rep protein sufficient for packaging, and
- an AAV cap protein sufficient for packaging;
- b) adding to the cell culture sodium propionate to a final concentration between about 0.5 mM and about 3 mM;
- c) adding to the cell culture sodium chloride in sufficient amount to increase the final concentration of the sodium chloride by between about 20 mM and 50 mM;
- and
- d) maintaining the cell culture under conditions that allow packaging to produce the rAAV particles,
- wherein the cell is transfected using a chemical based transfection method,
- wherein the sodium propionate and the sodium chloride are added after transfecting the cell, and
- wherein the sodium propionate and the sodium chloride are added no more than about 48 hours after transfecting the cell.
Dependencies are inferred from the claim text.