Microfluidic T-Chip enables one-step clinical-scale T-cell purification from blood products for CAR T-cell manufacturing

Rana, M.; Nigrovic, S. E.; Payan-Medina, A.; Saha, S.; Putaturo, V. R.; Cunneely, Q. E.; Bell, R.; Antmen, E.; Maus, M. V.; Toner, M.; Elsallab, M.; Mishra, A.

doi:10.64898/2026.07.30.741793

bioRxiv · 10.64898/2026.07.30.741793

Microfluidic T-Chip enables one-step clinical-scale T-cell purification from blood products for CAR T-cell manufacturing

Rana, M.·Nigrovic, S. E.·Payan-Medina, A.·Saha, S.·Putaturo, V. R.·Cunneely, Q. E.·Bell, R.·Antmen, E.·Maus, M. V.·Toner, M.·Elsallab, M.·Mishra, A.

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Abstract

Treatment with chimeric antigen receptor (CAR) T cells has emerged as a promising immune therapy for relapsed and refractory hematologic malignancies. The CAR T cells are manufactured in a series of steps that involve isolating T cells from the patients leukapheresis product, genetically modifying them to express the CAR against the target antigen, and reinfusing them into the patient. Efficient T-cell enrichment from leukapheresis products is critical to the success of these therapies. Current methods for T-cell sorting on a clinical scale involve several washing steps to remove red blood cells and platelets, followed by T-cell selection and activation. These multi-step processes result in cell loss during processing and involve several handling steps. Here, we utilize fluidically assembled micromagnetic lenses to develop a high-throughput, continuous-flow microfluidic T-cell sorter, designated as the T-Chip, for sorting magnetic bead-labeled CD3+ T cells in a single step. Our approach allows direct sorting of T cells in expansion media from leukopaks without any washing steps, effectively removing 99.999% of RBCs and platelets from the leukapheresis product. A single 1-inch x 3-inch T-Chip can process leukapheresis product at a throughput of 60 mL/hr and 2.56 {+/-} 0.12 billion cells/hr. Using this optimized workflow, we demonstrate clinical-scale enrichment of highly pure CD3+ T cells (97.7 {+/-} 1.3%) with high viability (97.0 {+/-} 1.1%) and recovery (87.3 {+/-} 14.8%) in a functionally closed manner. Downstream processing of T cells isolated using the T-Chip yielded potent anti-mesothelin CAR T cells with demonstrated anti-tumor efficacy. Overall, by exploiting precisely engineered magnetic forces and laminar flow, the microfluidic T-Chip overcomes bottlenecks caused by low throughput and enables single-step large-scale T-cell purification for the rapid development of CAR T cells.

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Rana, M., Nigrovic, S. E., Payan-Medina, A., Saha, S., Putaturo, V. R., Cunneely, Q. E., Bell, R., Antmen, E., Maus, M. V., Toner, M., Elsallab, M., Mishra, A.. 2026-07-31. Microfluidic T-Chip enables one-step clinical-scale T-cell purification from blood products for CAR T-cell manufacturing. https://doi.org/10.64898/2026.07.30.741793

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