The DOI matches its Crossref record; 99 of the 100 references it cites were found in a registry or an index.
Comparative analysis of assays to measure CAR T cell-mediated cytotoxicity
Open accessAbstract
The antitumor efficacy of genetically engineered 'living drugs', including chimeric antigen receptor and T-cell receptor T cells, is influenced by their activation, proliferation, inhibition, and exhaustion. A sensitive and reproducible cytotoxicity assay that collectively reflects these functions is an essential requirement for translation of these cellular therapeutic agents. Here, we compare various in vitro cytotoxicity assays (including chromium release, bioluminescence, impedance, and flow cytometry) with respect to their experimental setup, appropriate uses, advantages, and disadvantages, and measures to overcome their limitations. We also highlight the US Food and Drug Administration (FDA) directives for a potency assay for release of clinical cell therapy products. In addition, we discuss advanced assays of repeated antigen exposure and simultaneous testing of combinations of immune effector cells, immunomodulatory antibodies, and targets with variable antigen expression. This review article should help to equip investigators with the necessary knowledge to select appropriate cytotoxicity assays to test the efficacy of immunotherapeutic agents alone or in combination.
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“A U.S. company that seeks to manufacture CAR T-cells in the U.S. and E.U. must comply with both sets of regulations and work with strict oversight from both the FDA and the European Medicines Agency (EMA) to meet respective manufacturing standards (133,135,136).”
From Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy (2021) · cites this paper as [133]
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“A similar clinical trial phase structure exists in the European Union, although some GMP regulations are stricter in Europe (132,133).”
From Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy (2021) · cites this paper as [133]
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Cites
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Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy
Abou-el-Enein, M., Elsallab, M., Feldman, S. A., Fesnak, A. D., Heslop, H. E., Marks, P., Till, B. G., Bauer, G., & Savoldo, B. (2021) · Blood Cancer Discovery · doi:10.1158/2643-3230.bcd-21-0084 · Cited by 321
Deep generative models design mRNA sequences with enhanced translational capacity and stability
Zhang, H., Liu, H., Xu, Y., Huang, H., Liu, Y., Wang, J., Qin, Y., Wang, H., Ma, L., Xun, Z., et al. (2025) · Science · doi:10.1126/science.adr8470 · Cited by 98
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
GMP-Compliant Manufacturing of TRUCKs: CAR T Cells targeting GD2 and Releasing Inducible IL-18
Glienke, W., Dragon, A. C., Zimmermann, K., Martyniszyn, A., Mertens, M., Abken, H., Rössig, C., Altvater, B., Aleksandrova, K., Arseniev, L., et al. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.839783 · Cited by 79
Metadata
- Authors
- Kiesgen, S., Messinger, J. C., Chintala, N. K., Tano, Z., & Adusumilli, P. S.
- Affiliations
- Memorial Sloan Kettering Cancer Center (US)
- Corresponding author
- Adusumilli, Prasad S.
- Venue
- Nature Protocols
- Published
- 2021-02-15
- Volume
- 16
- Issue
- 3
- Pages
- 1331-1342
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- PMC
- PMC8064272
- OpenAlex
- W3132904428
- Cited by
- 138
- References
- 100
- Citations by year
- 2026: 24 · 2025: 37 · 2024: 32 · 2023: 22 · 2022: 18 · 2021: 5
- FWCI
- 2.33
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Cancer Institute (R01CA236615, R01CA235667, P30CA008748, T32CA009501)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Keywords
- chimeric antigen receptor T cells, T cell cytotoxicity, chromium release assay, bioluminescence assay, flow cytometry, potency assay, T cell exhaustion, T cell proliferation, combination immunotherapy, immunomodulatory antibodies, T cell receptor T cells
- MeSH terms
- Receptors, Chimeric Antigen, Animals, Cytotoxicity Tests, Immunologic, Flow Cytometry, Humans, Receptors, Antigen, T-Cell, T-Lymphocytes, Immunotherapy, Adoptive, Xenograft Model Antitumor Assays
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