The clinical translation of allogeneic natural killer cell therapies faces scalability challenges, including inconsistent NK cell expansion, variable product quality, and complex, labor-intensive manufacturing processes.

In a collaborative effort, we integrated CD3/28 Dynabead technology with the Dynacellect Magnetic Separation and Rotea Centrifugation platforms to revolutionize allogeneic NK manufacturing. On Day 0, a T-cell depletion is performed on peripheral blood mononuclear cells using the Active Release CD3/28 Dynabead technology on the Dynacellect Magnetic Cell Separation platform, which efficiently reduces residual T cells in the NK enrichment fraction. The Rotea Centrifugation system can wash the NK enriched fraction to NKXpander media with high yield, enabling a seamless transition into large-scale, feeder-free expansion in a single-use bioreactor. This process can achieve >95% NK cell purity by Day 21 and up to 900x expansion, with maintained cytotoxic potential.

Dynacellect further optimizes cell processing through an automated, closed-system design, reducing manual interventions and contamination risks while improving reproducibility. The physical and digital integration of Rotea and Dynacellect via Cellmation software streamlines workflows, enabling real-time monitoring and process control. This automation not only enhances manufacturing efficiency but also supports the production of "off-the-shelf" NK therapies, making these treatments more accessible to a broader patient population by reducing costs and production timelines.

Our approach demonstrates superior NK cell purity, robust expansion, and reduced processing times. This high-throughput, automated system ensures batch-to-batch consistency, meeting clinical scalability demands. Additionally, the closed-system design aligns with Good Manufacturing Practice (GMP) standards, simplifying regulatory compliance.

In conclusion, our automated end-to-end NK manufacturing workflow provides an innovative solution to overcome key scalability barriers in allo-NK therapy. This integrated platform enhances manufacturing efficiency, product consistency, and safety, advancing the clinical potential of NK cell-based immunotherapies and broadening patient accessibility to these transformative treatments.