Sunday, November 8, 2009 - 4:00 PM
S6
Production of a Secreted Recombinant Protein In Escherichia coli: Evolution of a Scalable Fermentation Process
Bruce F. Bishop, Sriram Srinivasan, Paul B. Lanter, and Will Miller. Global Biologics, Pfizer, 700 Chesterfield Village Parkway, Chesterfield, MO 63017
Advancement of therapeutic proteins to market requires the development of an efficient and robust production process capable of delivering consistent quality drug substance. In E. coli, recombinant therapeutic proteins may be expressed in a soluble or insoluble form in the cytoplasm, or secreted into the periplasmic space. High density fed-batch fermentation processes have long been shown to be viable strategies to produce these proteins at high levels using such expression routes. During the early stages of process development, it is possible to evaluate both intracellular and extracellular expression systems for product titer, product quality and suitability for downstream processing unit operations. Once an expression route is chosen, fermentation process development studies focus on media design, parameter evaluations and nutrient or substrate feeding strategies. A case study will be presented which describes the evaluation of cyptoplasmic inclusion body and periplasmic secretion routes of expression to produce a therapeutic protein in E. coli. Media development and glucose feeding strategy optimization, as related to process robustness for scale-up, will also be discussed.