Reducing Host Cell Burden By Strategic Genome Design

The design of Escherichia coli to improve bioprocessing is the subject of this presentation, a project which combines biochemical engineering, bioseparation, and proteomics.  Since downstream processing typically comprises a substantial portion of development and manufacturing, improvements in cell lines that reduce host cell burden is greatly desired.  Our efforts to improve ion exchange chromatography and immobilized metal affinity chromatography has demonstrated the fact that by understanding and manipulating the proteome of the host, improvements in purity and more importantly, column capacity can be realized.  During this presentation, a mathematical framework will be described that is used to guide host cell changes.  The design, construction, and deployment of this framework will be shown to be effective by various illustrative case studies of E. coli expression and purification.  Finally, we will demonstrate significant increases in column capacity – ion exchange in particular – and discuss the value of the approach to academia and industry.