IP Library › Granted Patent US 12,612,644
Granted Patent B2
US 12,612,644 · App. 17/267,247 · Granted Apr 28, 2026

Scalable method for recombinant AAV production

Inventors: Michael Gillmeister (Hanover, MD); Miguel Valle (Potomac, MD); Robert Stadelman (New Market, MD); Franz Gerner (Myersville, MD); Matthieu Guibert (Germantown, MD); Bhargavi Kondragunta (Clarksburg, MD)
Assignee: REGENXBIO INC.
C12N15/86C07K14/01C07K14/015C12N15/85C12N15/8645C12N2750/14151C12N2750/14152
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Quick Facts
Patent No.
US 12,612,644
App. No.
17/267,247
Granted
Apr 28, 2026
Kind
B2
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.

Claims (25)

1 . 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.

2 . The method of claim 1 , wherein prior to adding the sodium chloride, the cell culture comprises between about 90 mM and about 120 mM NaCl.

3 . The method of claim 1 , wherein the sodium chloride is added after adding the sodium propionate.

4 . The method of claim 1 , wherein the sodium chloride is added between about 5 minutes and about 6 hours after adding the sodium propionate.

5 . The method of claim 1 , wherein the sodium propionate is added between about 1 hour and about 48 hours after transfecting the cell.

6 . The method of claim 1 , wherein the cell is a HEK293 cell, HeLa cell, SF-9 cell, BHK cell, Vero cell, or PerC6 cell.

7 . The method of claim 1 , wherein the cell culture is a suspension culture.

8 . The method of claim 1 , further comprising recovering the rAAV particles.

9 . The method of claim 1 , wherein the cell culture produces between about 5×10e+10 genome copy (GC)/ml and about 1×10e+12 GC/ml rAAV particles.

10 . The method of claim 1 , wherein the cell culture produces at least about twice as many rAAV particles measured as GC/ml than a culture in sodium propionate chloride the absence of adding of the sodium propionate and the sodium chloride.

11 . The method of claim 1 , wherein the rAAV particles comprise a capsid protein of the AAV8, AAV9, AAV.rh10, AAV.rh20, AAV.rh39, AAV.Rh74, AAV.RHM4-1, or AAV.hu37 serotype.

12 . The method of claim 1 , wherein the rAAV genome comprises a transgene.

13 . The method of claim 1 , wherein the chemical based transfection method uses polyethylenimine (PEI).

14 . The method of claim 1 , wherein the sodium propionate and the sodium chloride are added separately in any order after transfecting the cell.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 12, 2021
From: GILLMEISTER, MICHAEL; VALLE, MIGUEL; STADELMAN, ROBERT; GERNER, FRANZ M.; GUIBERT, MATTHIEU; KONDRAGUNTA, BHARGAVI
To: REGENXBIO INC.
Reel/Frame 055240/0675 →
Continuity (2)
Provisional Application 62717212 · Aug 10, 2018
Related Publication 20210163991A1 · Jun 3, 2021
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