Real‐time dissolved carbon dioxide monitoring I: Application of a novel in situ sensor for CO2 monitoring and control

• Published in: Biotechnology and bioengineering Vol. 117; no. 4; pp. 981 - 991
• Main Authors: Chopda, Viki R., Holzberg, Timothy, Ge, Xudong, Folio, Brandon, Tolosa, Michael, Kostov, Yordan, Tolosa, Leah, Rao, Govind
• Format: Journal Article
• Language: English
• Published: New York Wiley Subscription Services, Inc 01-04-2020; John Wiley and Sons Inc
• Subjects: Baking yeast; bioprocess monitoring and control; Bioreactors; Calibration; Carbon dioxide; Cell culture; Diffusion rate; dissolved carbon dioxide; Flasks; Metabolism; mini bioreactor; Monitoring; Partial pressure; process analytical technology; Quality management; Sampling; Sensors; shake flask; Silicones; surface aeration intensification; Yeasts
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.27253

Dissolved carbon dioxide (dCO2) is a well‐known critical parameter in bioprocesses due to its significant impact on cell metabolism and on product quality attributes. Processes run at small‐scale faces many challenges due to limited options for modular sensors for online monitoring and control. Traditional sensors are bulky, costly, and invasive in nature and do not fit in small‐scale systems. In this study, we present the implementation of a novel, rate‐based technique for real‐time monitoring of dCO2 in bioprocesses. A silicone sampling probe that allows the diffusion of CO2 through its wall was inserted inside a shake flask/bioreactor and then flushed with air to remove the CO2 that had diffused into the probe from the culture broth (sensor was calibrated using air as zero‐point calibration). The gas inside the probe was then allowed to recirculate through gas‐impermeable tubing to a CO2 monitor. We have shown that by measuring the initial diffusion rate of CO2 into the sampling probe we were able to determine the partial pressure of the dCO2 in the culture. This technique can be readily automated, and measurements can be made in minutes. Demonstration experiments conducted with baker's yeast and Yarrowia lipolytica yeast cells in both shake flasks and mini bioreactors showed that it can monitor dCO2 in real‐time. Using the proposed sensor, we successfully implemented a dCO2‐based control scheme, which resulted in significant improvement in process performance.
A novel portable sensor for real‐time monitoring of dissolved CO2 (dCO2) in bioprocesses was developed. The measurement is based on the diffusion of dCO2 from the culture broth into a silicone sampling probe. The proposed sensor was successfully implemented in shake flasks and mini‐bioreactors for real‐time dCO2‐based control, which enables precise regulation of metabolic events in the bioprocesses and results in significant improvement in process performance.