P28: Titer and Recovery Yield Improvement of a Therapeutic Protein Produced in Escherichia coli by Reducing Protein Production Rate

Monday, November 7, 2011
Capri Ballroom (Marriott Marco Island)
Jianlin Xu, Yueming Qian, Paul M. Skonezny, Li You, Zizhuo Xing, David S. Meyers, Robert J. Stankavage and Zheng Jian Li, Cell Culture and Fermentation Process Development, Bristol-Myers Squibb, Syracuse, NY

A high protein production rate using a high inducer concentration at a high temperature is a general strategy for recombinant protein production in inclusion bodies (IBs) by E. coli, which usually includes applications of strong promoter such as T7 and isopropyl-b-D-thiogalactopyranoside (IPTG) induction at 37°C in a complex medium.  However, a new strategy that uses a slower protein production rate to improve recombinant protein titer and yield in IBs is reported in this study. This strategy using a lower feeding rate and a lower induction temperature at 29°C without adding any exogenous inducers increased the therapeutic protein (TP) volumetric titer by 84% and improved the specific production yield from 210 to 330 mg TP per g of dry cell weight. Furthermore, the new fermentation process significantly decreased an undesired variant, methionyl-TP, from 23.6% to 9.6%. The purified IB yield from the new process broth was also improved.

Keywords E. coli, therapeutic protein, inclusion body, fed batch fermentation, methionylation, and auto-induction

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