From Strain Selection To Purification – Key Factors For Successful Plasmid Production

By Stefan Schlößer, Eric Vincent, Ruchika Sharma, Samuel Ellis, Ines Hartmann, Eppendorf SE, Jülich, Germany, Eppendorf SE, Hamburg, Germany, Promega, Madison, WI USA, Thomson Instrument Company, Oceanside, CA USA

Plasmids serve as important tools in genetic engineering, gene therapy research or genetic vaccination. Recombinant plasmid DNA (pDNA) is produced in bacterial cultures, mostly in Escherichia coli (E. coli). Successful pDNA production, reaching high yield and quality, relies on multiple factors. These include the type of vector and host strain, the insert, and the methodology chosen for cultivation and downstream purification.

Review the distinct steps in the plasmid production workflow and give tips on optimization: from vector and strain selection to cultivation, from shake flasks to fermenters up to downstream processing steps of harvesting and purification.