Mononuclear cell derived NK cells
AssigneeImmunityBio, Inc.
InventorsRohit Duggal, Ranjeet Sinha, Wenzhao Li, Jason Isaacson, Karl Marquez, Patrick Soon-Shiong
Life of this patent
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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).
Claims16
A method of producing NK cells, comprising:
- isolating from whole blood or cord blood a mixture of mononuclear cells,
- contacting, in a container, the mixture of the mononuclear cells with an anti-CD16 antibody, an anti-CD3 antibody, and N-803 to activate NK cells;
- sequentially feeding, in the same container, the activated NK cells by adding to the container a medium containing N-803 to promote preferential growth of NK cells in the mixture until the activated NK cells constitute at least 90% 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 a mixture of mononuclear cells isolated from whole blood or cord blood, comprising:
- providing a mixture of the mononuclear cells that contains NK cells;
- contacting, in a container, the mixture of the mononuclear cells with an anti-CD16 antibody, an anti-CD3 antibody, and N-803 to activate NK cells;
- feeding, in the same container, the activated NK cells by adding to the container a medium containing N-803 to promote preferential growth of NK cells in the mixture until the activated NK cells constitute at least 90% 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, comprising:
- incubating a mixture of mononuclear cells isolated from whole blood or cord blood in an activation medium containing N-803, an anti-CD3 antibody, 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 in the same cell culture container by adding a medium containing N-803 to promote preferential growth of NK cells in the mixture, wherein the feeding is controlled by a predetermined schedule and/or a result from the step of measuring growth of the cells, and wherein the cells are fed until the NK cells are enriched to an at least 80-fold expansion;
- 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.