Abstract
Autologous CAR-T cell therapy has demonstrated remarkable clinical efficacy in hematologic malignancies, yet its broader application remains limited by complex, labor-intensive cell therapy manufacturing and inconsistent product quality. We describe a novel microfluidic cell separation platform based on deterministic lateral displacement (DLD), integrated into a fully automated, closed-system instrument (Curate system). N = 150 leukopacks were processed at a flow rate of 400 mL h(-1) with processed volumes up to 250 mL, white blood cell concentrations up to 168 M mL(-1) and total white blood cell counts up to 24 billion white blood cells. Compared to Ficoll®-based density gradient centrifugation, microfluidic DLD processing yielded significantly higher leukocyte recovery (88% vs. 58%), superior platelet and red blood cell depletion, and reduced CD69(+) T cell activation. Flow cytometric analysis revealed improved phenotypic preservation across key T cell subsets, including naïve and central memory populations. Cytokine profiling demonstrated enhanced washing efficiency, with markedly lower levels of biologic response modifiers. DLD-purified T cells exhibited enhanced expansion kinetics and greater yield, supporting improved manufacturing outcomes. These findings position microfluidic DLD-based processing as a clinically relevant, scalable alternative to conventional methods, with potential to improve consistency, potency, and accessibility of CAR-T therapies.