Abstract
Chimeric antigen receptor (CAR) T cell therapy that involves genetic engineering a patient’s own immune cells with antigen-specific receptors has shown remarkable efficacy in blood cancer treatment. Numerous clinical studies with CAR T cells targeting the blood cell surface protein CD19 led to the FDA ’s first approval of a genetically engineered cell therapy. The process of generating potent CAR T cells involves several carefully performed manufacturing steps. Here, we describe the generation of redirected engineered human CAR T cells for preclinical studies starting with the CAR design, retroviral gene transfer, detection of CAR expression, and expansion of transduced T cells.
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Abbreviations
- CAR:
-
Chimeric antigen receptor
- FACS:
-
Fluorescence-activated cell sorting
- FBS:
-
Fetal bovine serum
- GaLV :
-
Gibbon ape leukemia virus
- ICS:
-
Intracellular signaling
- IL:
-
Interleukin
- mAb:
-
Monoclonal antibody
- ORF :
-
Open reading frame
- RN :
-
RetroNectin
- RT :
-
Room temperature
- scFv :
-
Single chain variable fragment
- SN:
-
Supernatant
- SP :
-
Signal peptide
- TM:
-
Transmembrane domain
- VH:
-
IgG heavy chain variable region
- VL :
-
IgG light chain variable region
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Bunse, M., Höpken, U.E. (2022). Generation of Redirected Engineered Human Chimeric Antigen Receptor (CAR) T Cells. In: Walther, W. (eds) Gene Therapy of Cancer. Methods in Molecular Biology, vol 2521. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2441-8_4
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DOI: https://doi.org/10.1007/978-1-0716-2441-8_4
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