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Comparison of two lab-scale protocols for enhanced mRNA-based CAR-T cell generation and functionality
Nadine von Auw1, Robert Serfling1, Reni Kitte1
1Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), Perlickstr. 1, 04103, Leipzig, Germany.
Scientific Reports
|October 24, 2023
Summary
Optimizing media and T cell activators is crucial for CAR-T cell therapy manufacturing. This study evaluated two protocols for generating chimeric antigen receptor (CAR)-T cells to assess scalability and GMP compliance.
Area of Science:
- Cellular immunotherapy
- Biotechnology
- Process development
Background:
- Chimeric antigen receptor (CAR)-T cell therapies require robust manufacturing processes for clinical translation.
- Scaling up lab-based CAR-T cell production to automated Good Manufacturing Practice (GMP) environments presents significant challenges.
- Optimizing cell culture media and T cell activators is critical for maintaining cell viability, expansion, and functionality during CAR-T cell manufacturing.
Purpose of the Study:
- To compare two distinct protocols for generating CAR-mRNA-modified T cells at a lab scale.
- To analyze key parameters including cell viability, expansion, modification efficiency, and functionality.
- To evaluate the suitability of each protocol for upscaling and process development in mRNA-based CAR-T cell therapy manufacturing.
Main Methods:
- Two protocols for CAR-mRNA-modified T cell generation were compared using a standardized lab-scale process.
- Cell viability, expansion rates, CAR expression levels, and T cell functionality were assessed for each protocol.
- Parameters relevant to Good Manufacturing Practice (GMP) compliance and process scalability were analyzed.
Main Results:
- Protocol A demonstrated superior cell viability and expansion compared to Protocol B.
- Both protocols achieved comparable CAR-mRNA modification efficiency and T cell functionality.
- Specific medium and activator combinations showed significant impact on cell expansion and viability.
Conclusions:
- The selection of optimal medium and T cell activator is paramount for successful CAR-T cell therapy manufacturing.
- Protocol A shows greater potential for upscaling due to enhanced cell viability and expansion.
- Further process development focusing on GMP-compliant automation is recommended for mRNA-based CAR-T cell therapies.
