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Manufacturing and Analytical Characterization - Chemical

(M1430-11-64) System Robustness and Best Practices of a High Throughput LC-MS Method for Cell Culture Media Nutrient and Metabolite Analysis

Monday, October 21, 2024
2:30 PM – 3:30 PM MT
Purpose: Timely in-process monitoring of critical components in culture media is vital for optimal cell growth, quality, and yield in biotherapeutics process optimization. A 9 min high throughput LC-MS based method was developed and tested for the detection and quantification of cell culture media nutrient and metabolites. This work describes subjecting the spent media analysis method for multi-week, long term data studies and on multiple instruments, capturing robustness, accuracy and precision metrics.  This performance data demonstrates the method’s suitability for daily monitoring of in-process samples in upstream development environment.   

Methods: Spent media samples from cell culture experiment expressing mAb protein were used. LC-MS instrument used was a BioAccord™ HRMS system using an ACQUITY™ Premier LC system. Waters_connect™ was used for data acquisition and processing. Additional details can be found in Waters application notes. 

Results: A recently developed LC-MS method having acquisition time of 9 min has made it possible to perform daily monitoring of culture media and metabolites in a timely basis.  Rapid data collection and processing enables bioprocess engineers to analyze the data and make adjustment decisions on daily basis, if needed. To ensure system robustness under such conditions, the method was tested assuming daily sampling, performing spent media analyses intertwined with intact protein (high level glycoprofiling) analyses. Studies were performed using two BioAccord systems in two Waters locations in two different counties. The data was processed in terms of reproducibility of response, retention time, mass error, and other attributes of all compound’s class observed in the spent media.  Stability monitoring of standard LC-MS calibration solutions in a two-week cell culture product production cycle was also monitored. Result suggests calibration data collected at day 0 can be used throughout the two weeks incubation data processing.  

Conclusion: A robust and practical LC-HRMS methodology supports the routing monitoring of nutrient and metabolite and intact mass glycoprofiling assisting upstream bioprocessing development.