Monday, November 9, 2009
P18
Pichia pastoris therapeutic protein production; process development and understanding
Jeffrey Bernstein, Bioprocess Research and Development, Merck & Co., RY80Y-120, 126 E. Lincoln Ave, Rahway, NJ 07065
We show a number of strategies used to characterize and enhance the production of therapeutic proteins in the yeast Pichia pastoris. Molecular profiling tools such as microarray and qPCR are used to monitor global transcription and RNA abundance of target production genes and their regulatory elements. Spectrophotometric assays determine the level of protease activity in different production strains. FACS analysis is employed to monitor cell viability during shake flask and bioreactor stages, defining ranges which maximize cell robustness and is also used to monitor target products as well as glycan characteristics. These techniques are used in conjunction with more traditional tools such as spent medium analysis and sensors to measure residual methanol during induction. Coupled with fermentation strategies that seek to control process parameters, these methods support optimal production of target proteins.
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