Process integration and intensification are an indispensable part of current biopharmaceutical manufacturing practices to overcome the production and quality-related issues and ongoing stiff market competition over consistent product stability. Integration of upstream and downstream unit operations is the key to achieving a true continuous process with minimal hold steps, shorter residence times, and desired product attributes. Acoustic wave separation (AWS) using three-dimensional standing acoustic waves is the next generation continuous clarification technique that has potential merits over existing membrane-based devices for high cell density perfusion cultures. In this study, Cadence<sup>TM</sup> Acoustic Separator (CAS) was used as the cell retention technique for Chinese Hamster Ovary (CHO) cells operated in perfusion culture. The effect of critical process parameters (CPPs) on the cell separation efficiency and protein recovery, such as perfusate flowrate, concentrate flowrate, acoustic power and load cell density, were evaluated in fed-batch studies. Based on the results, a hybrid model was developed using both physics-based and data-driven predictions to estimate optimum clarification efficiency. A distributed control system was integrated with AWS, coupling the hybrid model with a control algorithm to monitor deviations in incoming load cell density and implement proportionate control compared to the model setpoint. Perfusion runs were performed with cultivations up to 15 days, and separation efficiencies greater than 80% were achieved while facing no issue of fouling or pressure drop. The three-dimensional acoustophoresis resulted in higher levels of cell concentrations, viability and fewer clumps. The hybrid control strategy demonstrated in this study endorses the potential of AWS as an efficient ‘filterless’ technology to achieve enhanced continuous clarification and can be used for seamless upstream-downstream integration.

Acoustic wave separator as a cell retention device for integrated CHO cell perfusion process

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.