Aqueous Two‐Phase Extraction of Monoclonal Antibodies from High Cell Density Cell Culture

Improvements of the upstream process for the production of monoclonal antibodies (mAb) has resulted in mammalian cell cultures with significantly increased cell densities. These high cell density (HCD) broths pose severe challenges for established downstream unit operations due to the increased biomass load. To meet these challenges, aqueous two‐phase extraction was examined as a potential method for the clarification, as well as first stage purification of a Chinese hamster ovary expressed mAb with cell densities up to 70 · 106 cells mL−1. A clarification with 99 % yield of mAb and 68 % DNA removal was accomplished, which contributes to enable a foundation to the industrial application of aqueous two‐phase extraction for manufacturing of mAb from HCD cultivations. High cell density approaches in the upstream process offer great potential for an intensified production of monoclonal antibodies (mAb). However, significant challenges are imposed on the subsequent downstream process. In this study, proof of concept for clarification, capture and first purification of mAb from high cell density cell cultures by aqueous two‐phase extraction is presented.