Raman upstream bioprocess control

Nicro combined fermentation system engineering with Endress+Hauser Raman technology and analytical services from BICRO BIOCentre to create a complete solution for fermentation processes. Inline monitoring replaces time-consuming offline analysis with real-time measurements of metabolites and biomass. This gives scientists greater process insight, supports timely intervention, reduces contamination risk, and enables optimized production and increased yield.

The original cost analysis estimated a return on investment after 35 fermentation processes when inline Raman monitoring was compared with offline sample testing. The analysis also determined that the faster turnaround of Raman-monitored fermentation could increase plant productivity by 33% over a 10-day period.

Real-time insight improves fermentation control

Direct inline measurement of glycerol as an input metabolite, acetate as a byproduct, and OD600 for biomass growth provides deeper insight into fermentation performance. By monitoring multiple attributes through a single probe, the solution enables process decisions to be based on current conditions rather than delayed laboratory results. Results include:

• Faster process cycle times through real-time metabolite monitoring
• Improved process control supporting optimized production and increased yield
• Reduced contamination risk by limiting offline sample handling
• Simultaneous monitoring of multiple fermentation parameters
• Reduced time between batches
• A complete packaged technology solution for fermentation processes

Fermentation progress can be estimated by monitoring dissolved oxygen in the media. Optical density and metabolite concentrations provide additional insight, but these parameters have traditionally been measured offline using time-consuming chromatographic techniques.

Removing samples for analysis delays access to process information and introduces additional handling. It can also create a risk of contamination and the potential loss of a valuable batch. Nicro needed a solution that would provide timely information about metabolite concentration and biomass growth without relying on repeated offline sampling.

Nicro combined its fermentation engineering expertise with BICRO BIOCentre and Endress+Hauser technologies and services to provide a complete packaged solution for fermentation processes. The solution includes:

• Engineering and design of a complete fermentation system from Nicro
• An Endress+Hauser Raman Rxn analyzer
• An Endress+Hauser Raman Rxn-45 probe
• Analytical services from BICRO BIOCentre
• S-PACT modeling services supporting analytical model development

The Raman probe is installed directly inline and monitors glycerol concentration, acetate concentration, and OD600 for biomass growth. Multiple attributes can be monitored simultaneously through one probe, providing a real-time view of fermentation conditions and eliminating the delay associated with offline sample analysis.

Inline information also helps scientists recognize process events that may not occur at exactly the same point in every batch. In the documented application, online monitoring allowed scientists to identify when bacteria switched from one carbon source to another, supporting intervention at the appropriate feed point.