Enabling Fully Closed CAR-T Cell Manufacturing with ProPak™ GMP Liquid Cytokines

Comparing Closed and Open CAR-T Cell Manufacturing Workflows

As CAR-T cell therapies advance toward clinical production, manufacturers are increasingly focused on simplifying and standardizing their workflows. Traditional protocols relying on lyophilized cytokines often involve time-consuming reconstitution and open handling steps that increase cleanroom demands and contamination risk. R&D Systems™ ProPak GMP liquid cytokines, supplied in ready-to-use, weldable, single-use bags, are designed to seamlessly integrate into fully closed-system manufacturing processes.

This application note provides a direct comparison of ProPak liquid cytokines used in closed-system workflows and conventional lyophilized cytokines used in open systems. The results demonstrate that ProPak liquid IL-7 and IL-15 support robust CAR-T cell expansion, maintain critical T cell memory phenotypes, and preserve functional potency, matching the performance of lyophilized formats. These findings confirm that ProPak liquid cytokines enable efficient scaling of CAR-T cell production from research to clinical manufacturing without compromising cell quality or therapeutic efficacy.

Key Takeaways

• Ready-to-use ProPak GMP liquid cytokines packaged in process-sized, single-use bags demonstrate full recovery of IL-7 and IL-15 concentrations in GMP Human T Cell Media, enabling streamlined cytokine handling in closed systems.
• Both research-scale, open and clinical-scale, fully closed CAR-T cell manufacturing workflows support robust T cell expansion (>30-fold) and viability (>95%), with the closed system producing significantly higher total viable CAR-T cell yields at clinical scale.
• The closed and open workflows maintain favorable T cell memory phenotypes, with >60% of CD4⁺ and CD8⁺ T cells exhibiting combined stem cell memory (T_SCM) or central memory (T_CM) markers, alongside low PD-1 expression, indicating minimal exhaustion.
• Functional potency assays confirm that CAR-T cells generated in both workflows exhibit equivalent, target-specific killing of CD19⁺ cells and secrete key effector cytokines including IFN-γ, Granzyme B, IL-2, and TNF-α at comparable levels.
• These results demonstrate that transitioning to a fully closed, ProPak-enabled CAR-T cell manufacturing process maintains critical cellular characteristics and functional efficacy, while simplifying handling and improving scalability.
   

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