Implementation of Design of Experiment (DoE) for process development and optimization of biosimilars

Abstract

Selection of a suitable basal media, feed, feeding strategy and glycan structures has a pivotal role during the process development and optimization of monoclonal antibodies. Instead of using one-factor-at-time approach or classical approach for media and feed selection, a DoE approach was implemented. A Simplex centroid mixture design was applied to identify the suitable media/media combinations by evaluating cell growth and productivity followed by a full factorial design for screening of the best suitable feed that enhanced the productivity. Optimization of the process was performed using Response surface methodology with central composite design to determine the optimum interaction between the selected media and feeds. Statistically designed experiments were performed to determine the factors influencing glycan. Uridine, MnCl2 and galactose were added as per the factorial design for evaluating their effect on modulating glycosylation. Prediction of the best combination was made with the maximum response titer by Design-Expert software 12.0. A threefold increase in titer (1909 mg/L) and significant improvement in glycosylation were obtained by statistical approach.