Immunophenotyping of CAR T cell fitness using flow cytometry

MBTP 60 = Miltenyi Biotec-tested panel 60

This application protocol describes the determination of the cell fitness of chimeric antigen receptor (CAR) T cell products using Annexin V and 7-AAD for discrimination between viable, apoptotic, and dead cells. To demonstrate this state-of-the-art panel for CAR T cell fitness, CAR T cells were generated within 12 days using the CliniMACS Prodigy® T Cell Transduction process. 

Protocol

Antibody panel for CAR T cell fitness

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
7-AAD*-Viability-655-730 (blue)
Annexin VREA1165ApoptosisFITC525/50 (blue)
CD4*REA623Helper T cellsVioGreen™525/50 (violet)
CD8*REA734Cytotoxic T cellsAPC-Vio® 770750 LP (red)
CD3REA613T cellsAPC655-730 (red)
CD19 CAR FMC63REA1297CAR+ cellsPE585/40 (blue)

* Alternative CE-IVD/ASR reagents available.

Gating strategy

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Materials

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