Assessment of CAR T cell frequencies in axicabtagene ciloleucel and tisagenlecleucel patients using duplex quantitative PCR
Chimeric antigen receptor (CAR) T cell (CART) therapy has been established as a treatment option for patients with CD19-positive lymphoid malignancies in both the refractory and the relapsed setting. Displaying significant responses in clinical trials, two second-generation CART products directed ag...
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| Hauptverfasser: | , , , , , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
30 September 2020
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| In: |
Cancers
Year: 2020, Jahrgang: 12, Heft: 10 |
| ISSN: | 2072-6694 |
| DOI: | 10.3390/cancers12102820 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3390/cancers12102820 Verlag, lizenzpflichtig, Volltext: https://www.mdpi.com/2072-6694/12/10/2820 
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| Verfasserangaben: | Maria-Luisa Schubert, Alexander Kunz, Anita Schmitt, Brigitte Neuber, Lei Wang, Angela Hückelhoven-Krauss, Sascha Langner, Birgit Michels, Antje Wick, Volker Daniel, Carsten Müller-Tidow, Peter Dreger, and Michael Schmitt |
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| 245 | 1 | 0 | |a Assessment of CAR T cell frequencies in axicabtagene ciloleucel and tisagenlecleucel patients using duplex quantitative PCR |c Maria-Luisa Schubert, Alexander Kunz, Anita Schmitt, Brigitte Neuber, Lei Wang, Angela Hückelhoven-Krauss, Sascha Langner, Birgit Michels, Antje Wick, Volker Daniel, Carsten Müller-Tidow, Peter Dreger, and Michael Schmitt |
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| 520 | |a Chimeric antigen receptor (CAR) T cell (CART) therapy has been established as a treatment option for patients with CD19-positive lymphoid malignancies in both the refractory and the relapsed setting. Displaying significant responses in clinical trials, two second-generation CART products directed against CD19, axicabtagene ciloleucel (axi-cel) and tisagenlecleucel (tisa-cel), have been approved and integrated into the clinical routine. However, experimental assay for quantitative monitoring of both of these CART products in treated patients in the open domain are lacking. To address this issue, we established and validated a quantitative single copy gene (SCG)-based duplex (DP)-PCR assay (SCG-DP-PCR) to quantify CARTs based on the FMC63 single chain variable fragment (scFv), i.e., axi-cel and tisa-cel. This quantitative PCR (qPCR) approach operates without standard curves or calibrator samples, offers a tool to assess cellular kinetics of FMC63 CARTs and allows direct comparison of CART-copies in axi-cel versus tisa-cel patient samples. For treating physicians, SCG-DP-PCR is an important tool to monitor CARTs and guide clinical decisions regarding CART effects in respective patients. | ||
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