(675f) Advancing Real-Time Live Cell Imaging Technologies to Support the Development of Downstream Purification Processes for Therapeutic Viruses
AIChE Annual Meeting
Thursday, November 17, 2022 - 5:15pm to 5:36pm
Adenovirus samples with a GFP-transgene were initially used, with the IncucyteÂ® Live-Cell Analysis System and the IncucyteÂ® AI Confluence software implemented to optimise cell culture regimes and cell confluence over time to ensure assay viability. Segmentation algorithms provided by the IncucyteÂ® Base Analysis software were used to estimate virus titer values by the appearance of fluorescently labelled cells, which were compared to estimates obtained through traditional Hexon staining. Subsequently, adenovirus samples without a GFP transgene were used to extend the Incucyteâs ability to provide label-free titer estimates using machine learning methodologies, with the results similarly validated using Hexon staining and compared. The comparative costs and labour were then tabulated, demonstrating the relative benefits of the Incucyte® based assay as the number of samples increases.
Overall, investigation of optimized virus purification techniques is essential as specialized biotherapeutics gain clinical prominence. However, development of analytical assays that can match increased purification throughput has lagged, making sample analysis the bottleneck in virus therapeutic production. The assay presented in this work has been specifically developed to handle the sampling requirements and virus titer range associated with virus membrane chromatography, demonstrating the potential of Incucyte® integrated live-cell analysis techniques in the analytical workflow to enable new separation processes.