Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes
Abstract Chimeric Antigen Receptor (CAR) T cell therapy is a pivotal treatment for hematological malignancies. However, CAR T cell products exhibit batch-to-batch variability in cell number, quality, and in vivo efficacy due to donor-to-donor heterogeneity, and pre/post-manufacturing processes, and...
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Nature Portfolio
2025-05-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-59789-w |
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| author | Kerwin Kwek Zeming Kai Yun Quek Wei-Xiang Sin Denise Bei Lin Teo Ka-Wai Cheung Chin Ren Goh Faris Kairi Elizabeth Lee Francesca Lorraine Wei Inng Lim Michaela Su-Fern Seng Shui Yen Soh Michael E. Birnbaum Jongyoon Han |
| author_facet | Kerwin Kwek Zeming Kai Yun Quek Wei-Xiang Sin Denise Bei Lin Teo Ka-Wai Cheung Chin Ren Goh Faris Kairi Elizabeth Lee Francesca Lorraine Wei Inng Lim Michaela Su-Fern Seng Shui Yen Soh Michael E. Birnbaum Jongyoon Han |
| author_sort | Kerwin Kwek Zeming |
| collection | DOAJ |
| description | Abstract Chimeric Antigen Receptor (CAR) T cell therapy is a pivotal treatment for hematological malignancies. However, CAR T cell products exhibit batch-to-batch variability in cell number, quality, and in vivo efficacy due to donor-to-donor heterogeneity, and pre/post-manufacturing processes, and the manufacturing of such products necessitates careful testing, both post-manufacturing and pre-infusion. Here, we introduce the Cell Trajectory Modulation (CTM) assay, a microfluidic, label-free approach for the rapid evaluation of the functional attributes of CAR T cells based on biophysical features (i.e., size, deformability). CTM assay correlates with phenotypic metrics, including CD4:CD8 ratio, memory subtypes, and cytotoxic activity. Validated across multiple donors and culture platforms, the CTM assay requires fewer than 10,000 cells and delivers results within 10 minutes. Compared to labeled flow cytometry processing, the CTM assay offers real-time data to guide adaptive manufacturing workflows. Thus, the CTM assay offers an improvement over existing phenotypic assessments, marking a step forward in advancing CAR T cell therapy manufacturing. |
| format | Article |
| id | doaj-art-ce7048909bbe4d4f9d4020d7ac69060d |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-ce7048909bbe4d4f9d4020d7ac69060d2025-08-20T02:33:32ZengNature PortfolioNature Communications2041-17232025-05-0116111510.1038/s41467-025-59789-wCell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypesKerwin Kwek Zeming0Kai Yun Quek1Wei-Xiang Sin2Denise Bei Lin Teo3Ka-Wai Cheung4Chin Ren Goh5Faris Kairi6Elizabeth Lee7Francesca Lorraine Wei Inng Lim8Michaela Su-Fern Seng9Shui Yen Soh10Michael E. Birnbaum11Jongyoon Han12Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Department of Haematology, Singapore General HospitalSingHealth Duke-NUS Oncology Academic Clinical Programme, Sing Health Duke-NUS Academic Medical CentreSingHealth Duke-NUS Oncology Academic Clinical Programme, Sing Health Duke-NUS Academic Medical CentreCritical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Critical Analytics for Manufacturing of Personalized Medicine, Singapore-MIT Alliance for Research and Technology (SMART)Abstract Chimeric Antigen Receptor (CAR) T cell therapy is a pivotal treatment for hematological malignancies. However, CAR T cell products exhibit batch-to-batch variability in cell number, quality, and in vivo efficacy due to donor-to-donor heterogeneity, and pre/post-manufacturing processes, and the manufacturing of such products necessitates careful testing, both post-manufacturing and pre-infusion. Here, we introduce the Cell Trajectory Modulation (CTM) assay, a microfluidic, label-free approach for the rapid evaluation of the functional attributes of CAR T cells based on biophysical features (i.e., size, deformability). CTM assay correlates with phenotypic metrics, including CD4:CD8 ratio, memory subtypes, and cytotoxic activity. Validated across multiple donors and culture platforms, the CTM assay requires fewer than 10,000 cells and delivers results within 10 minutes. Compared to labeled flow cytometry processing, the CTM assay offers real-time data to guide adaptive manufacturing workflows. Thus, the CTM assay offers an improvement over existing phenotypic assessments, marking a step forward in advancing CAR T cell therapy manufacturing.https://doi.org/10.1038/s41467-025-59789-w |
| spellingShingle | Kerwin Kwek Zeming Kai Yun Quek Wei-Xiang Sin Denise Bei Lin Teo Ka-Wai Cheung Chin Ren Goh Faris Kairi Elizabeth Lee Francesca Lorraine Wei Inng Lim Michaela Su-Fern Seng Shui Yen Soh Michael E. Birnbaum Jongyoon Han Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes Nature Communications |
| title | Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes |
| title_full | Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes |
| title_fullStr | Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes |
| title_full_unstemmed | Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes |
| title_short | Cell trajectory modulation: rapid microfluidic biophysical profiling of CAR T cell functional phenotypes |
| title_sort | cell trajectory modulation rapid microfluidic biophysical profiling of car t cell functional phenotypes |
| url | https://doi.org/10.1038/s41467-025-59789-w |
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