Closed-system manufacturing of CD19 and dual-targeted CD20/19 chimeric antigen receptor T cells using the CliniMACS Prodigy device at an academic medical center
Abstract
Background aims Multiple steps are required to produce chimeric antigen receptor (CAR)-T cells, involving subset enrichment or depletion, activation, gene transduction and expansion. Open processing steps that increase risk of contamination and production failure are required. This complex process requires skilled personnel and costly clean-room facilities and infrastructure. Simplified, reproducible CAR-T-cell manufacturing with reduced labor intensity within a closed-system is highly desirable for increased availability for patients. Methods The CliniMACS Prodigy with TCT process software and the TS520 tubing set that allows closed-system processing for cell enrichment, transduction, washing and expansion was used. We used MACS-CD4 and CD8-MicroBeads for enrichment, TransAct CD3/CD28 reagent for activation, lentiviral CD8 TM-41BB-CD3 ζ-cfrag vectors expressing scFv for CD19 or CD20/CD19 antigens for transduction, TexMACS medium-3%-HS-IL2 for culture and phosphate-buffered saline/ethylenediaminetetraacetic acid buffer for washing. Processing time was 13 days. Results Enrichment (N = 7) resulted in CD4/CD8 purity of 98 ± 4.0%, 55 ± 6% recovery and CD3 + T-cell purity of 89 ± 10%. Vectors at multiplicity of infection 5-10 resulted in transduction averaging 37%. An average 30-fold expansion of 10 8 CD4/CD8-enriched cells resulted in sufficient transduced T cells for clinical use. CAR-T cells were 82-100% CD3 + with a mix of CD4 + and CD8 + cells that primarily expressed an effe…
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Further analysis of T cell subsets revealed substantial proportions of TSCM and TCM in the majority of cell products.
“Further analysis of T cell subsets revealed substantial proportions of TSCM and TCM, which are early memory T cells possessing long-term persistence and antitumor activity in vivo, in the majority of cell products, which is consistent with previous studies using a similar platform and reagents.14,19,25”
From Point-of-care manufacturing of anti-CD19 CAR-T cells using a closed production platform: Experiences of an academic in Thailand (2024) · cites this paper as [25]
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Promises and challenges of a decentralized CAR T-cell manufacturing model
Shah, M., Krull, A., Odonnell, L., de Lima, M. J., & Bezerra, E. (2023) · Frontiers in Transplantation · doi:10.3389/frtra.2023.1238535 · Cited by 77 · see citing works
Luanpitpong, S., Klaihmon, P., Janan, M., Kungwankiattichai, S., Owattanapanich, W., Kunacheewa, C., Chanthateyanonth, S., Donsakul, N., U-pratya, Y., Warindpong, T., et al. (2024) · Molecular Therapy: Oncology · doi:10.1016/j.omton.2024.200889 · Cited by 17 · see citing works
- Authors
- Zhu, F., Shah, N., Xu, H., Schneider, D., Orentas, R., Dropulic, B., Hari, P., & Keever-Taylor, C. A.
- Affiliations
- Medical College of Wisconsin (US)
- Lentigen Technology (United States) (US)
- Corresponding author
- Keever-Taylor, Carolyn A.
- Venue
- Cytotherapy
- Publisher
- Elsevier BV
- Published
- 2018-03
- Volume
- 20
- Issue
- 3
- Pages
- 394-406
- ISSN
- 1465-3249 (ISSN-L)
- OpenAlex
- W2781388493
- Cited by
- 120
- References
- 4
- Citations by year
- 2026: 7 · 2025: 13 · 2024: 13 · 2023: 18 · 2022: 14 · 2021: 20 · 2020: 14 · 2019: 16 · 2018: 3 · 2017: 1
- FWCI
- 4.10
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Cytotherapy (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- CAR T cell manufacturing, CD19, chimeric antigen receptor T cells, CliniMACS Prodigy, lentiviral transduction, T cell enrichment, T cell activation, T cell expansion, IFN-gamma production, cytotoxicity, academic medical centers
- MeSH terms
- Academic Medical Centers, Receptors, Chimeric Antigen, B-Lymphocytes, Cell Line, Cytological Techniques, Humans, Receptors, Antigen, T-Cell, T-Lymphocytes, T-Lymphocytes, Cytotoxic, Transduction, Genetic, CD4-Positive T-Lymphocytes, Immunophenotyping, CD28 Antigens, CD8-Positive T-Lymphocytes, Antigens, CD19, Antigens, CD20
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