High throughput downstream process development for cell based products using aqueous two phase systems.
As the clinical development of cell based therapeutics has evolved immensely within the past years, downstream processing strategies become more relevant than ever. Aqueous two phase systems (ATPS) enable the label free, scalable, and cost effective separation of cells, making them a promising tool for downstream processing of cell based therapeutics. Here, we report the development of an automated robotic screening that enables high throughput cell partitioning analysis in ATPS. We demonstrate that this setup enables fast and systematic investigation of factors influencing cell partitioning. Moreover, we examined and optimized separation conditions for the differentiable promyelocytic cell line HL 60 and used a counter current distribution model to investigate optimal separation conditions for a multi stage purification process. Finally, we show that the separation of CD11b positive and CD11b negative HL 60 cells is possible after partial DMSO mediated differentiation towards the granulocytic lineage. The modeling data indicate that complete peak separation is possible with 30 transfers, and >93% of CD11b positive HL 60 cells can be recovered with >99% purity. The here described screening platform facilitates faster, cheaper, and more directed downstream process development for cell based therapeutics and presents a powerful tool for translational research.
Journal of chromatography. A