Mononuclear cell derived NK cells
AssigneeImmunityBio, Inc.
InventorsRohit Duggal, Ranjeet Sinha, Wenzhao Li, Jason Isaacson, Karl Marquez, Patrick Soon-Shiong
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Abstract
Cord blood or peripheral blood NK cells are prepared from whole blood mononuclear cells without the need to isolate CD34+ hematopoietic stem cells or NK cells, and without the need for a feeder layer. Advantageously, the methods presented herein use an enrichment process that uses antiCD16 agonist antibodies, antiCD3 antibodies, and N-803. Moreover, contemplated processes are suitable for adaptation into a fully automated production process (GMP in a box).
Claims20
A method of expanding NK cells from a mixture of mononuclear cells, the method comprising:
- providing in a container a mixture of the mononuclear cells that contains equal or less than 5% NK cells;
- contacting the mixture of the mononuclear cells with an anti-CD16 antibody and N-803 to activate NK cells;
- and
- feeding in the same container the activated NK cells with a medium containing N-803 until the activated NK cells constitute at least about 80% of all live cells or until the activated NK cells are enriched to an at least 80-fold expansion.
A method of expanding NK cells in an automated bioreactor, the method comprising:
- incubating a mixture of mononuclear cells in an activation medium containing N-803 and an anti-CD16 antibody for a time sufficient to activate NK cells;
- wherein the mixture of mononuclear cells is contained in a cell culture container while incubating the mixture;
- measuring growth of the cells while the cells are in the container;
- automatically feeding the cells using a medium containing N-803 until the activated NK cells constitute at least about 80% of all live cells or until the activated NK cells are enriched to an at least 80-fold expansion, wherein the feeding is controlled by a predetermined schedule and/or a result from the step of measuring growth of the cells;
- terminating feeding the cells wherein the terminating is controlled by a predetermined schedule and/or a result from the step of measuring growth of the cells.
Dependencies are inferred from the claim text.