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US 12,180,457 B2Granted
Closed-system manufacturing process for CAR-T cells
AssigneeSeattle Children's Hospital
InventorsMichael C. Jensen, Joshua Gustafson
Life of this patent
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Abstract
Some embodiments provided herein relate to methods and compositions for making genetically modified T cells. In some such embodiments, CD4+ and CD8+ T cells are cultured in a single serum-free volume. In some embodiments, co-cultured CD4+ and CD8+ T cells can be transduced with a lentiviral vector, and a population of transduced T cells can be harvested within a shorter period of time than other conventional methods.
Claims12
A method of manufacturing genetically modified T cells, wherein the genetically modified T cells comprise a chimeric antigen receptor (CAR), the method comprising:
- (a) providing CD8+ T cells and CD4+ T cells in a culture media in a single culture vessel, wherein the culture media comprises IL-7, IL-15, and IL-21, and lacks serum, thereby obtaining co-cultured T cells;
- (b) initiating the CD8+ T cells and CD4+ T cells for transduction, thereby obtaining initiated T cells, wherein the initiation is performed for a period within a range from about 12 hours to about 24 hours;
- (c) transducing the initiated T cells with a vector encoding the CAR, thereby obtaining transduced T cells, wherein the transducing comprises:
- (i) magnetically concentrating the initiated T cells, thereby obtaining concentrated T cells, and
- (ii) incubating the concentrated T cells with a transduction media for 4 hours or less;
- and
- (d) measuring an amount of the transduced T cells, wherein the amount is at least sufficient for a therapeutic dose of the transduced T cells in an adoptive cell transfer to a human subject, and harvesting the transduced T cells, thereby obtaining the genetically modified T cells, wherein step (d) is performed less than 7 days or less after step (b) is performed.
4 more elements
Dependencies are inferred from the claim text.