Biologics and AAV-based gene therapies are gaining more market share for disease treatment. However, developing a robust and efficient mammalian cell line for their production is challenging and usually requires extensive cellular characterization. Historically, a flow cytometer is utilized in these cell-based assays. However, a flow cytometer is relatively expensive and involves extensive training for both operation and maintenance. Recently, with an increase in computing capabilities and high-quality camera sensors, image-based cytometry has been innovated to provide a precise and cost-effective alternative for cell line process development. In this work, we described a cell line development workflow incorporating an image-based cytometer, namely the Countstar Rigel, for transfection efficiency assessment and stable pool evaluation using CHO and HEK293 cells expressing antibody and rAAV vector, respectively. In the two case studies, we demonstrated:
1. Countstar Rigel provided similar detection accuracy to flow cytometry.
2. Countstar Rigel -based pool evaluation can help determine the desirable group for single-cell cloning (SCC).
3. Countstar Rigel incorporated cell line development platform achieved 2.5 g/L mAb titer.
We also discussed the possibility of using Countstar as another layer of an rAAV DoE-based optimization target.
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