P5: Automated On-line Sampling and Nutrient Monitoring for Characterizing Glucose Consumption of Microbial Fermentation Cultures

A Seg-Flow® automated on-line vessel sampling system (Seg-Flow System) was evaluated for characterizing real-time glucose consumption of microbial fermentation cultures used for protein biotherapeutic production.  High cell density fermentation processes employing host strains of Scarab Genomics’ Clean Genome® E. coli were conducted at 10L working volume using a biphasic approach.  Cultures were grown to a specific density in minimal salts/glucose media during the batch phase.  Upon reaching the targeted density, glucose was fed exponentially to control the culture growth rate at a specified constant µ hr^-1during the fed-batch phase.  The Seg-Flow System, consisting of the Seg-Flow autosampling module, a YSI® Biochemistry Analyzer and a fraction collector, provided real-time analysis of media glucose concentrations as well as programmed time-point sampling for manual glucose analysis and additional off-line analytics.  Automated and manual glucose analyses were comparable, which demonstrated fidelity of the results generated by the Seg-Flow System.  The Seg-Flow System provided an automated, real-time solution for monitoring glucose concentrations and, in turn, rapidly characterizing glucose consumption of E. coli fermentation cultures.