Anoushka Durve, Dharani Vora, Jeff Goby, Eike Zimmermann and Kenji Furuya
During the development of therapeutic monoclonal antibodies (mAbs), monitoring protein productivity along with product quality (PQ) attributes is essential. There is limited understanding on how PQ profiles change with process conditions or culture duration. Real-time monitoring capabilities would allow us to understand such changes during upstream processing. At Boehringer Ingelheim, Fremont, we have already established the Waters PATROL™ UPLC to measure real-time daily titers during culture duration. Product quality testing, however, requires samples to be affinity purified, prohibiting the use of the PATROL™ UPLC directly for permeate analysis, for e.g. size variants by SEC or charge variants by CEX. We thus have enhanced the PATROL™ system to a 2D platform with protein A purification in the 1st dimension followed by a PQ assay in the 2nd dimension. This system allows real-time analysis of PQ attributes such as size or charge variants directly during cell culture samples, without the time-consuming affinity purification operation. In this talk, we have demonstrated a proof of concept to utilize the 2D PATROL™ UPLC to monitor PQ attributes during cell culture processing.
Take home message-
The impact of process conditions and culture duration on mAb size and charge variant profiles can be monitored in real-time during continuous cell culture processing using the enhanced 2D PATROL™ UPLC.