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A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
JoVE 杂志
免疫与感染
This content is Free Access.
JoVE 杂志 免疫与感染
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells

A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells

DOI:
10.3791/59033-v

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08:46 min

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November 12, 2019

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, , , , , , , , , , , , ,
1ACEA Biosciences, Inc, 2ProMab Biotechnologies. Inc, 3ACEA Therapeutics

Chapters

  • 00:04Title
  • 00:30Chimeric Antigen Receptor (CAR)-Encoding Lentivirus Generation
  • 02:29CAR T Cell Generation and Expansion
  • 03:16CAR Expression Detection
  • 04:55Real-Time Cytolysis Potency Assay
  • 06:48Results: Representative CAR T Cell Target Cell Killing Efficacy as Assessed by RTCA
  • 08:05Conclusion

Summary

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We describe a quantitative real-time in vitro cytolysis assay system to evaluate the potency of chimeric antigen receptor T cells targeting liquid and solid tumor cells. This protocol can be extended to assess other immune effector cells, as well as combination treatments.

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Real-time Potency AssayChimeric Antigen Receptor T CellsSolid And Hematological Cancer CellsCELLigence Real-time Cell AnalysisLabel-free MonitoringTarget Cancer Cell KillingEffector Cell TypesCombination TherapiesLentiviral VectorHEK293FT CellsTransfectionLentivirus ProductionUltracentrifugationQuantitative RT-PCRCAR-T Cell GenerationPeripheral Blood Mononuclear CellsCD3/CD28 Coated Microbeads

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