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Related Experiment Video

Updated: Jan 4, 2026

Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
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Immunophenotypic Analysis of CAR-T Cells.

Júlia Teixeira Cottas de Azevedo1,2,3, Amanda Mizukami4, Pablo Diego Moço4

  • 1Center for Cell-based Therapy, Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil. juliatca@usp.br.

Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2019
PubMed
Summary

Chimeric antigen receptor T-cell (CAR-T) immunotherapy shows promise for cancer treatment. Characterizing CAR-T cell phenotype during bioprocessing using flow cytometry is vital for quality control and enhancing anti-tumor efficacy.

Keywords:
CAR-T cellsCentral memory T cellEffector T cellEffector memory T cellFlow cytometryImmunophenotypic analysisMemory stem T cellsNaive T cellT cells subsets

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Area of Science:

  • Immunology
  • Cellular Therapy
  • Biotechnology

Background:

  • CAR-T cell immunotherapy is a significant advancement in cancer treatment.
  • The efficacy of CAR-T cell therapy depends on the phenotype, activation, and function of the infused cells.
  • Consistent quality control of CAR-T cells during manufacturing is essential for therapeutic success.

Purpose of the Study:

  • To propose a flow cytometry panel for comprehensive immunophenotypic characterization of CAR-T cells.
  • To establish a standardized method for assessing CAR-T cell subsets during bioprocessing.
  • To ensure the quality and potential efficacy of CAR-T cell products.

Main Methods:

  • Development and application of a multi-parameter flow cytometry panel.
  • Characterization of distinct CAR-T cell subsets based on immunophenotypic markers.
  • Analysis of CAR-T cells throughout the bioprocessing workflow.

Main Results:

  • The proposed flow cytometry panel effectively distinguishes and characterizes various CAR-T cell subsets.
  • Detailed immunophenotypic profiles of CAR-T cells were obtained.
  • The method provides critical data for CAR-T cell quality control.

Conclusions:

  • Immunophenotypic characterization of CAR-T cells via flow cytometry is crucial for bioprocess monitoring.
  • The developed panel aids in ensuring the quality and potential efficacy of CAR-T cell therapies.
  • Standardized characterization contributes to improved patient outcomes in CAR-T cell immunotherapy.