(567bw) Efficient and Effective Supplement Screening for the Development of Chemically Defined Media in Mammalian Cell Culture

Development and refinement of chemically defined media (CDM) is of principal importance towards upstream process development due to regulatory concerns and the desire for process robustness. Typically, media is comprised of a mixture of complex and defined components. Frequently, the supplementation/removal of any one of these components has nominal effects on the resulting cell culture performance, other times a major effect is observed. We have developed a streamlined and reliable screening method to screen supplements in a high-throughput format which may be beneficial for both early-stage and late-stage upstream projects. Performance in our milliliter scale cell culture system was correlated via cell growth and titer. Fractional factorial designs were employed to optimize media blending on an individual compound basis. Thousands of media designs tested hundreds of media compounds over the course of only a few months, and a statistically optimized CDM was developed which facilitated a 123% increase in peak viable cell density and a 103% increase in harvest titer. This increase in performance was subsequently verified in multiple cells lines expressing multiple antibodies in addition to other CDM medias with beneficial results. Use of the improved CDM in laboratory scale bioreactors was also verified to have an improvement in culture performance, with comparable product quality. Further multivariate data analysis revealed unexpected correlations between various compounds in our statistical analysis. The compound-by-compound screening approach highlighted in this work has provided a robust, scalable, and economical method for inclusion into future media development and refinement strategies.