Process Improvements for Production of a Complex Glycosylated Fusion Protein which Delivers a Two-Fold Increase in Product Titre and Comparable Product Quality

摘要

This work describes the improvement of a high yielding commercial process of a complex glycosylated fusion protein, resulting in almost a two-fold increase in product titre while maintaining comparable product quality to the established process. The improvement in titre is achieved by adjusting a number of process parameter set-points, resulting in higher cell densities and higher titres in the production culture. In order to sustain these higher cell densities, the production culture is supplementedwith a nutrient-rich tailored feed medium. During the development of this higher-yielding process, the resultant doubling of titre was accompanied by increased levels of mis-folded and aggregated product. In order to overcome this phenomenon, low molecular weight medium additives (Peptide X and Copper) were added to the production culture, resulting in modulation of mis-folded and aggregated product to levels comparable to the control. All product quality attributes including N-linked oligosaccharide sialylated species in the yield enhanced process are maintained to comparable control levels. This work demonstrates how process improvements can be introduced to facilitate a two-fold increase in process yield, while simultaneously maintaining the required product quality attributes.