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Updated: Jul 1, 2026
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Manufacturing Chimeric Antigen Receptor (CAR) T Cells for Adoptive Immunotherapy
Published on: December 17, 2019
Standardized Flow Cytometry Assay Using Dried Recombinant Antibody Panels for CAR T Cell Characterization
Annika Ridzal1, Anja Brauchle2, Desiré Missing2
1Miltenyi Biotec; annikag@miltenyi.com.
Journal of Visualized Experiments : Jove
|June 29, 2026
Summary
Dried recombinant antibody panels streamline Chimeric Antigen Receptor T (CAR T) cell analysis via flow cytometry. This ready-to-use format reduces preparation steps and operator variability for more consistent critical quality attribute assessment.
Area of Science:
- Immunology
- Biotechnology
- Analytical Chemistry
Background:
- Flow cytometry is essential for characterizing Chimeric Antigen Receptor T (CAR T) cells, assessing critical quality attributes like viability, phenotype, and transduction efficiency.
- Conventional liquid reagent workflows involve multiple preparation steps, leading to potential variability and operator-dependent inconsistencies.
- Recombinant antibody technology offers enhanced specificity and reduced lot-to-lot variability compared to traditional antibodies.
Purpose of the Study:
- To present a streamlined, step-by-step workflow for CAR T cell analysis using preformulated dried recombinant antibody panels.
- To demonstrate the practical application of dried antibody panels in multiparametric CAR T cell characterization.
- To compare the hands-on time and efficiency of dried reagent panels against conventional liquid reagent workflows.
Main Methods:
- Development and validation of a protocol for CAR T cell staining using dried recombinant antibody panels.
- Flow cytometry data acquisition using the optimized staining protocol.
- Analysis of CAR T cell critical quality attributes, including viability, phenotype, and transduction efficiency.
- Comparison of hands-on time and workflow efficiency with traditional liquid reagent methods.
Main Results:
- A practical, step-by-step workflow for CAR T cell analysis using dried recombinant antibody panels was established.
- Dried panels simplified reagent handling and reduced preparation steps, leading to decreased hands-on time.
- Representative staining data demonstrated the utility of dried panels for multiparametric CAR T cell characterization.
- Reduced lot-to-lot variability was observed due to the use of recombinant antibody technology.
Conclusions:
- Dried recombinant antibody panels offer a practical and efficient alternative to conventional liquid reagents for CAR T cell analysis.
- This approach enhances consistency and reduces operator variability in flow cytometry workflows.
- The presented framework facilitates the implementation of dried antibody panels for streamlined multiparametric characterization of CAR T cells.
