Abstract
Biomanufacturing processes may be optimized by storing cell culture media at room temperature, but this is currently limited by their instability and change in color upon long-term storage. This study demonstrates that one of the critical contributing factors toward media browning is tryptophan. LC-MS technology was utilized to identify tryptophan degradation products, which are likely formed primarily from oxidation reactions. Several of the identified compounds were shown to contribute significantly to color in solutions but also to exhibit toxicity against CHO cells. A cell-culture-compatible antioxidant, a-ketoglutaric acid, was found to be an efficient cell culture media additive for stabilizing components against degradation, inhibiting the browning of media formulations, and decreasing ammonia production, thus providing a viable method for developing room-temperature stable cell culture media.
Highlights
A cell-culture-compatible antioxidant, a-ketoglutaric acid, was found to be an efficient cell culture media additive for stabilizing components against degradation, inhibiting the browning of media formulations, and decreasing ammonia production, providing a viable method for developing room-temperature stable cell culture media
These combinations of reactions often yielded products with fully oxidized heterocycles and a high level of conjugation, which are typical properties in colored organic compounds, and correlates with the color increase in the solutions. As these compounds are significantly different in structure from Trp and as protein translation involves many quality control systems to avoid the incorporation of incorrect amino media acids (AAs) [43], it was deemed unlikely that the incorporation of these degradation products in the drug product would be a concern
The discovery that some of these compounds can contribute directly to CHO toxicity and feed browning indicates that these degradation processes can be detrimental in biomanufacturing, and methods need to be established to limit such degradation in cell culture media (CCM) that are intended to be stored at elevated temperatures
Summary
A cell-culture-compatible antioxidant, a-ketoglutaric acid, was found to be an efficient cell culture media additive for stabilizing components against degradation, inhibiting the browning of media formulations, and decreasing ammonia production, providing a viable method for developing room-temperature stable cell culture media. The latest drive in the industry is to improve the chemical stability of hydrated media against degrading conditions such as exposure to light and increased temperatures. CCM that are stable at room temperature (RT) may decrease the energy consumption by eliminating the need for refrigeration This optimization of CCM stability involves both developing methods to detect critical changes to CCM that can negatively impact cell culture [3,4] and using this information to discover strategies to mitigate such changes. When exposed to stressed conditions (e.g., light, heat), sensitive CCM components decompose and new degradation products are generated. This chemical change in the media composition can often be directly observed by a change in the color of the media
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