Development of cell culture processes to deliver proteins with optimal product quality attributes, while maintaining process robustness and ease of manufacturing, can be challenging. We present one such case study here, in which we simultaneously optimized the cell culture process for drug substance color, charge variants and glycation while making the process more robust and operationally simpler. Drug substance color is an important product quality attribute that is especially relevant for high concentration drug substance formulations. We achieved lower drug substance color by lowering concentrations of B vitamins and iron, adding hydrocortisone, substituting cystine for cysteine in the media and lowering culture temperature. We hypothesize that the oxidation of the antibody leads to increased antibody color. We used in vitro incubation experiments to screen over 50 compounds for antioxidant activity and identified two antioxidants capable of reducing antibody coloration. These novel antioxidants were then tested in cell culture studies and were found to reduce antibody coloration without impacting culture productivity. Changes to cell culture media aimed at lowering protein color led to lower growth which affected other product quality attributes such as C-Terminal lysine levels and glycation. Several media and process parameters were further optimized to reduce C-Terminal lysine.  In order to optimize glycation, we developed and employed a mathematical model to simulate the growth of cells, substrate consumption, protein synthesis and glycation. We used this model to develop a glucose feeding strategy that could deliver low and consistent levels of glycation.

Insights into the impact of cell culture media components on drug substance color, charge variants and glycation

247th National Meeting (2014)

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