CELL INTELLIGENCE · 肿瘤细胞治疗研究

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 patient's 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.