Automated CAR-T Cell Manufacturing: from Research to Point of Care production.

Background: Point-of-care manufacturing of CAR-T cells could reduce cost and improve patient access, but few closed, automated systems support both flexible process development and GMP production. Methods and results: We describe LimCORE, a single-use consumable for cell incubation and processing, implemented in LimGROW (manual system) and LimONE (automated system). In LimGROW, T cells expanded 246 +/-18-fold over 10 days at optimized seeding density, versus 89 +/-9-fold in a G-Rex control, with similar viability (>90%) and phenotype. Process parameters directly transferred to LimONE, where automated, closed buoyancy-based CD3+ selection achieved 90.3 +/-3.5% purity and 59.2 +/-17.0% recovery in under two hours. Using this workflow, we manufactured CD19 CAR-T cells in an automated 7-day LimONE process requiring 155 minutes of operator intervention. LimONE manufactured CAR-T cells reached 42.6 +/-2.4-fold expansion (532 +/-29 M cells at harvest), with 43.6 +/-6.0% transduction efficiency and viability similar to G-Rex controls. CD4+/CD8+ ratios, CD8+ differentiation subsets, and cytotoxicity were comparable between platforms, while LimONE-manufactured cells showed increased metabolic fitness. Across CAR-T manufacturing runs on the LimONE, no hardware or software failures occurred, and automated liquid transfers and volume concentration stayed within +/-5% accuracy. Conclusions: These data show that LimCORE supports T cell expansion, selection, and complete CD19 CAR-T manufacturing with performance similar to existing platforms while reducing operator time, across R&D and GMP scale. Key words: Automation; CAR-T; Point of care; T cell manufacturing