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Data Spotlight

Linking Fluorescence and mAb Yield for Clone Selection in CHO Cells

Background

Developing antibody-based oral vaccines requires large-scale production of monoclonal antibodies (mAbs) for in vivo studies. Outsourcing production can be costly, so many researchers turn to in-house cell culture systems to generate the necessary quantities. A key challenge in this process is the lengthy timeline—often six months or more—to identify stable, high-producing cell lines.

To address this bottleneck, researchers have leveraged a T2A peptide–based system linking antibody production with GFP expression. Because GFP and mAb are produced in equimolar amounts, intracellular GFP serves as a direct proxy for secreted antibody levels. By monitoring GFP expression, scientists can quickly select for high-producing, stable clones and track expression stability over time. When combined with temperature shift strategies in CHO-S cell culture, this approach enables higher yields and more efficient production cycles.

Results

Data Spotlight - Fluorescence Monitoring and mAb in CHO Cell Cultures

 

Materials & Methods

CHO-S cells stably secreting GFP-tagged monoclonal antibodies were cultured in 250 mL flat-bottom glass Erlenmeyer flasks with a 40% filling volume. Cultures were inoculated at a starting density of 3.0 × 10⁵ cells/mL and maintained at 37 °C with agitation at 150 rpm using a 25 mm orbital shaker diameter.

Conclusion

Using real-time monitoring of GFP expression and cell growth, researchers were able to validate that fluorescence reliably reflects antibody production, consistent with prior published findings. This proof-of-concept demonstrates how fluorescence can serve as a powerful, non-invasive marker for optimizing cell line selection and determining the optimal harvest point in antibody production workflows. By simplifying clone screening and production monitoring, the approach accelerates timelines, reduces costs, and strengthens in-house capabilities for therapeutic antibody development.

This research was conducted by:

Hans Van der Weken

Ghent University

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