Recycling and Regeneration of Spent Perfusion Media via Ion Concentration Polarization

Abstract

The widespread adoption of monoclonal antibody therapies is often constrained by their high prices, which can limit accessibility, particularly for patients in low- and middle-income countries. Addressing this economic barrier is crucial to ensure that life-saving treatments can reach all who need them. We present a series of bioprocessing innovations designed to reduce the cost of monoclonal antibody manufacturing and improve global access to these critical therapeutics. The work focuses on developing technologies for media regeneration and recycling, with the goal of reducing the economic and environmental impact of cell culture media in perfusion mammalian cell culture. We demonstrate a microfluidic separation device engineered to selectively remove metabolic waste products—specifically ammonia and lactate—from spent media using ion concentration polarization. Building on this foundation, a scalable millifluidic system was developed to enable higher-throughput waste removal. We characterized the impact of media recycling upon batch and perfusion cell cultures. We devised a nutrient supplementation strategy to create ‘regenerated’ media that minimized any effect on cell growth and productivity compared to fresh media. To support continuous manufacturing, a perfusion culture system incorporating a microfluidic spiral cell retention device and continuous cell bleed was established, and stable performance was maintained over extended durations. A further innovation introduced a multi-stage waste recovery system that increased media regeneration yield to 87.5%. This recovery rate enabled a self-recycling perfusion bioreactor in which 75% of the media feed was regenerated, without significant impact on cell growth, productivity, or product quality. Together, these advances establish a novel biomanufacturing platform that combines electrokinetic waste removal with media regeneration and recycling. The approach is broadly adaptable to mammalian cell culture processes and offers a promising path toward more sustainable, cost-effective, and environmentally responsible production of monoclonal antibodies and other biologics.