Monday, November 9, 2009 - 10:05 AM
S11

Optimization of an Escherichia coli Fermentation Process

Jane Gunson, Kate L. Moiseff, Ryan Hamilton, and Michael W. Laird. Late Stage Cell Culture - Process Research and Development, Genentech, Inc., 1 DNA Way, MS 32, South San Francisco, CA 94080

Optimization of an existing Escherichia coli fermentation process was achieved by the implementation of a general fed-batch platform fermentation process with in-process nutrient feeds. A two-fold increase in cell mass and product yield were observed with the improved fermentation process while maintaining comparable product quality to the existing process. Experiments to determine acceptable operating ranges for fermentation control parameters were initially performed under univariate conditions. The results indicated several parameters including pH and temperature increased the level of a product quality attribute when controlled outside the proposed target ranges. Acceptable operating ranges determined in the univariate experiments were applied to a 6-factor multivariate study designed to identify control parameter interactions impacting fermentation process performance and product quality.  The multivariate study was modified when it was observed that the acceptable operating ranges determined in the univariate study did not yield acceptable fermentation process performance or levels of a particular quality attribute. Narrower ranges for all parameters were required to successfully execute the 6-factor study plan. The results from these fermentations were used in combination with other representative development data in a risk ranking and filtering (RRF) exercise to determine which parameters impact process performance, product quality and process-related impurities. The output of this exercise provided a basis for a process characterization study design. An overview of the improved fermentation process and results from the univariate and multivariate studies will be presented. Furthermore, the output of the RRF exercise and its contribution to the fermentation process characterization design will also be discussed.       

See more of Microbial culture scale up, manufacturing, and process characterization
See more of Invited Oral Papers

See more of RAFT VIII (November 8 - 11, 2009)