Optimizing Cell Therapy Analytics Through Design Of Experiment Methodologies

Developing and commercializing cell therapies requires robust, phase-appropriate analytical assays to ensure product safety, consistency, and efficacy. Among these, potency assays, which directly measure a product’s biological activity, are especially challenging to design and validate. Complex cell-based functional assays often demand extensive time and resources to achieve required accuracy, sensitivity, and reproducibility, particularly as regulatory expectations increase through development stages. Applying Design of Experiment (DoE) methodologies streamlines assay optimization by evaluating multiple variables and interactions simultaneously, accelerating timelines and conserving clinical material.

In a recent case study, IFN-γ secretion assays, which assess T cell activation, were refined using DoE to identify critical factors such as effector-to-target ratio and incubation time, while demonstrating non-significant effects from media type and IL-2 concentration. Similarly, flow cytometry-based cell proliferation assays benefited from DoE, defining optimal parameters including T:E ratio, seeding density, dye pulse conditions, and co-culture duration. This structured, statistically driven approach enables the development of scalable, reproducible assays aligned with regulatory standards, supporting informed decision-making across discovery, clinical development, and manufacturing.