Process mapping and optimization study of CHO cell cultures for mAb production using Ambr® 250 high-throughput parallel bioreactors.

Abstract

The demand to accelerate monoclonal antibody (mAbs) process development timelines using Chinese hamster ovary (CHO) host cells to bring therapies to patients sooner is gaining momentum. The applicability of single use high-throughput (HTP) bioreactor system such as Ambr^®250 facilitating precise and automated control is very promising. This entails optimizing process parameters through design of experiments (DoE) using less resources and time, compared to traditionally employed large-scale bench top reactors (2-5L). It is imperative to improve mAb productivity through robust process development to mitigate current manufacturing challenges. In this study, a systematic mapping approach was employed to identify critical process parameters (CPP) and improve process efficacy. A central composite design (CCD) was used in Ambr^®250 bioreactors to investigate the impact of initial seeding density (SD) and feeding rate (FR) on mAb production. Variance in the SD and FR impacted the cell performance and mAb titer profile based on which parameter optimization was done using response surface methodology. Significant impact of FR and SD was identified leading to improved mAb titer. Bioreactors operated at SD > 1 × 10⁶ cells/mL and FR of > 2% Vc/day were more productive, and respective optimal FR and SD were estimated at 2.68% Vc/day and 1.1 × 10⁶ cells/mL. Cell viability and productivity were well-maintained at optimal conditions allowing extended cultivation time to reach higher mAb titer of up to 5 g/L. These findings, which optimize the operating range of critical process parameters (CPPs) using the high-throughput Ambr® 250 scaled-down platform, provide a framework for accelerated early-phase process development and enable reliable scalability to commercial manufacturing. Improving productivity and providing robust estimates for manufacturing scale would significantly cut costs and reduce timelines for biologics development and facilitate patient access.