Sunday, November 8, 2009 - 3:00 PM
S4
Investigating the use of Clean Genome® E. coli hosts for large scale cGMP manufacturing of pDNA drugs
Jesse D. McCool1, Maureen A. Hamilton1, Wenglong R. Lin1, Rayford J. Anderson1, Chih-Cheng Chen1, Jeff Gagnon1, Aaron Grinstein1, Tammy Lambert1, Reuben Land1, Amy M. Ledoux1, Seann T. O'Connell1, Scott Rose1, Christopher J. Dale1, Dogan Ornek1, Brad G. Sepp1, Catharine A. Street1, and Frederick R. Blattner2. (1) Microbial R&D Services, Lonza Biologics, Inc., 99 South Street, Hopkinton, MA 01748, (2) Scarab Genomics, LLC, 1202 Ann Street, Madison, WI 53713
There is growing interest in the development of safe and effective plasmid DNA based vaccines for a variety of human and veterinary applications. This has led to a multitude of recent studies focused on ways to improve aspects of pDNA manufacturing. An exquisite new technology from Scarab Genomics, L.L.C. offers, for the first time, a reduced genome E. coli host that permits the production of plasmid DNA in the complete absence of IS elements (Clean Genome® E. coli). In addition to other deleted genetic content such as pathogenicity islands and phage remnants, reduced genome hosts are expected to facilitate the development of improved processes, lower manufacturing costs and increase product safety in unprecedented ways. Here we report results of an on-going evaluation of one particular Scarab host (MDS42recA) in the context of microbial process development. Fed-batch fermentation results show that despite the absence of nearly 700,000 bp, MDSrecA performs better than a conventional host in terms of pDNA productivity and product quality attributes. One of the processes tested was scaled previously to 30L in our cGMP facilities.