Cellular and metabolic actuators to control monoclonal antibody β-1,4 galactosylation

The biopharmaceutical industry, a cornerstone of the global economy, witnessed monoclonal antibodies (mAbs) generating a staggering $217 billion in revenue in 2021 (80.2% of total biopharmaceutical product sales). Notably, amid the spotlight on COVID-19 vaccine research and production, over 90 approved mAbs continue to address critical diseases such as cancer, immune disorders, infectious diseases, and genetic disorders. With an annual rate of nearly 4 new mAb products gaining approval, their economic significance and life-saving potential have become increasingly apparent. This study aims to establish robust methods for producing mAbs with optimal and consistent galactosylation profiles. Two strategies were devised and implemented. The first strategy focused on gaining transcriptional control over the β1,4-galactosyltransferase gene through a synthetic gene circuit. The second strategy involved a metabolic engineering approach, employing dose-dependent galactosylation inhibition in a hypergalactosylated mAb. In conclusion, this research generates insights into optimizing mAb galactosylation profiles through innovative strategies. These findings have broad implications for enhancing product quality and aid product and process development in mAb manufacturing.