Effective Feeding Strategy to Enhance Cell Productivity in HEK293.2sus Fed-batch Cultures

Human embryonic kidney 293 (HEK293) are widely used for recombinant therapeutic protein production due to their stable growth and high protein expression capabilities. This study aimed to determine an optimal feeding strategy to maximize HEK293.2sus cell productivity in fed-batch cultures. We first investigated the growth curve of HEK293.2sus batch culture in BalanCD HEK293 medium to identify the most suitable initial feeding time, which was determined to be around day 3. Subsequently, six different intermittent feeding strategies were evaluated to optimize cell density and viability. The results demonstrated that intermittent feeding with 5% v/v feeding medium at the 3^rd and 5^th day of cultivation yielded the highest maximum viable cell density (15.59 × 10⁶ cells/mL) and viability (81.23%). This strategy requires less time and feeding medium while maintaining high cell concentration and productivity. These findings suggest an efficient and cost-effective approach for large-scale protein production using HEK293.2sus cells in fed-batch cultures.