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

Point-of-care manufacturing of CAR-T cells has the potential to lower costs and enhance patient access, yet limited closed, automated systems exist that can handle both flexible process development and GMP production. Researchers have developed LimCORE, a single-use consumable for cell incubation and processing, which can be implemented in both a manual system, LimGROW, and a fully automated, closed system, LimONE.

In LimGROW, T cells expanded 246% over 10 days with optimized seeding density, compared to 89% expansion in a control system, while maintaining 90% viability and similar phenotype. The parameters used in LimGROW were successfully transferred to LimONE, achieving 90.3% purity and 59.2% recovery of CD3+ cells within two hours through automated buoyancy-based selection.

Using this streamlined workflow, LimONE produced CD19 CAR-T cells in a fully closed, automated 7-day process, requiring only 155 minutes of operator intervention. The manufactured CAR-T cells exhibited an expansion factor of 42.6, with 43.6% transduction efficiency and comparable viability to control cells. Furthermore, the ratios of CD4+/CD8+ cells, CD8+ differentiation subsets, and cytotoxicity were consistent between platforms.

LimONE-manufactured cells also demonstrated increased spare respiratory capacity and oxygen consumption. Throughout four manufacturing runs on LimONE, no hardware or software failures were reported, and automated liquid transfers and volume concentration maintained accuracy within +/-5%.

These findings indicate that LimCORE can support T cell expansion, selection, and complete CD19 CAR-T manufacturing with performance comparable to existing platforms, while simultaneously reducing operator time across both R&D and GMP scale closed-system formats.