Immunophenotyping of stem cell-like memory T (Tscm) cells in CAR T cell products using flow cytometry

MBTP 45 = Miltenyi Biotec-tested panel 45

This application protocol describes the analysis of stem cell-like memory T (Tscm) cell frequencies in chimeric antigen receptor (CAR) T cell products. To demonstrate the state-of-the-art panel, CAR T cells were generated within 12 days using the CliniMACS Prodigy® T Cell Transduction process and stained for Tscm cells. The data was analyzed using Miltenyi Biotec’s automated data acquisition and analysis Express Mode program “CAR T Cell Express Mode Package”. 

Protocol

Antibody panel for Tscm cell analysis

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
7-AAD*-Viability-655-730 (blue)
CD3*REA613T cellsFITC525/50 (blue)
CD4*REA623Helper T cellsVioGreen™525/50 (violet)
CD8*REA734Cytotoxic T cellsAPC-Vio® 770750 LP (red)
CD45ROREA611T cell differentiationAPC655-730 (red)
CD197 (CCR7)*REA546 T cell differentiationVioBlue®450/50 (violet)
CD95 (FAS)REA738 T cell differentiationPE585/40 (blue)
CD62L*145/15T cell differentiationPE-Vio 770750 LP (blue)

* Alternative CE-IVD/ASR reagents available. 

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Materials

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