Enhancing biomanufacturing efficiency: A model-based plug-and-play Hybrid fed-batch process using ATF perfusion for high-yield drug substance production

The biopharmaceutical industry is witnessing a rapid growth, with the therapeutic antibody market expanding significantly due to the increasing demand for targeted therapies and personalized medicine. Traditional fed-batch processes for monoclonal antibody (mAb) production, while effective and well established, necessitate the use of large bioreactors and extensive supporting infrastructure. These requirements translate to high operational costs and complex logistics, which pose significant challenges for small and medium-sized enterprises (SMEs) that may lack the resources and capital to invest in such expansive setups. This study proposes a new single-step concentrated fed-batch process leveraging Alternating Tangential Flow (ATF) perfusion of the inoculum integrated to the production bioreactor to achieve high initial cell densities. A model-based approach was adopted to rationally optimise the process parameters. By integrating high seeding densities and optimised process strategies, this approach enhanced bioreactor efficiency and product yield by 4 to 6-fold (conventional 1.9 g/L to 10 - 12 g/L in intensified process), without compromising protein quality. The intensified process was further validated using different territory CHO cell lines and was found to yield similar results. An analysis of conventional fed-batch process to the new plug-and-play hybrid process suggested a 60 % reduction in costs, improvement in production efficiency, and consistent product quality, including post-translational modifications and glycosylation profiles. This scalable method is adaptable to various cell lines and biopharmaceutical products, offering a promising alternative to conventional methods and enabling faster, more cost-effective biomanufacturing.