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High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

STAR Protocols, ISSN: 2666-1667, Vol: 3, Issue: 1, Page: 101174
2022
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Article Description

Here, we present a comprehensive protocol for the generation and functional characterization of chimeric antigen receptor (CAR) T cells and their products by mass cytometry in a reproducible and scalable manner. We describe the production of CAR T cells from human peripheral blood mononuclear cells. We then detail a three-step staining protocol with metal-labeled antibodies and the subsequent mass cytometry analysis. This protocol allows simultaneous characterization of CAR T cell intracellular signaling, activation, proliferation, cytokine production, and phenotype in a single assay.

Bibliographic Details

Michelozzi, Ilaria M; Sufi, Jahangir; Adejumo, Thomas A; Amrolia, Persis J; Tape, Christopher J; Giustacchini, Alice

Elsevier BV

Neuroscience; Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology

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