Improved t cell manufacturing process
AssigneeCelgene Corporation
InventorsThomas A. BRIEVA, David HSIUNG, Seth JONES, Shiv MISTRY, Nayyereh RAJAEI
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Abstract
Provided herein are improved methods of manufacturing cells, including T cells and CAR T cells. Also provided herein are methods of manufacturing cells, such as T cells and CAR T cells, obtainable from blood using a method incorporating membrane filtration and Ammonium-Chloride-Potassium (ACK) buffer to isolate the cells from other blood components.
Claims9
A method of manufacturing chimeric antigen receptor (CAR)-expressing T cells (CAR T cells) from peripheral blood mononuclear cells (PBMCs) from a subject from which a blood sample is obtained, comprising the steps of:
- a. obtaining PBMCs from the blood sample;
- b. isolating the PBMCs that have been obtained from the blood sample using a membrane filtration system and Ammonium-Chloride-Potassium (ACK) buffer;
- c. washing the PBMCs via centrifugation;
- d. optionally cryopreserving the PBMCs;
- e. optionally thawing the PBMCs cryopreserved in step (d);
- f. washing the PBMCs using a membrane filtration system;
- g. manufacturing the CAR T cells from the PBMCs from step (f);
- and
- h. washing the CAR T cells from step (g) using a membrane filtration system.
A method of manufacturing chimeric antigen receptor (CAR)-expressing T cells (CAR T cells) from peripheral blood mononuclear cells (PBMCs) from a leukapheresed blood sample from a subject, comprising the steps of:
- a. isolating the PBMCs from the leukapheresed blood sample using a membrane filtration system and Ammonium-Chloride-Potassium (ACK) buffer;
- b. cryopreserving the PBMCs from step (a);
- c. thawing the PBMCs from step (b);
- d. washing the thawed PBMCs from step (c) using a spinning filtration system;
- e. manufacturing the CAR T cells from the PBMCs from step (d);
- and
- f. washing the CAR T cells from step (e) using a membrane filtration system.
A method of manufacturing chimeric antigen receptor (CAR)-expressing T cells (CAR T cells) from peripheral blood mononuclear cells (PBMCs) from a blood sample from a subject, comprising the steps of:
- a. isolating the PBMCs from the blood sample using a membrane filtration system and Ammonium-Chloride-Potassium (ACK) buffer;
- b. cryopreserving and thawing the PBMCs from step (a);
- c. washing the thawed PBMCs from step (b) using a membrane filtration system;
- d. manufacturing the CAR T cells from the PBMCs;
- and
- e. washing of the CAR T cells using a spinning membrane filtration system.
Dependencies are inferred from the claim text.