From Single-Cell Analysis to Micro-Volume Whole Blood: How AbbVie Scientists extended the CETSA® Utility. 

How do you differentiate between a compound binding in a target cell versus a bystander cell? How do you measure target engagement in a clinical sample when you only have a few microliters of blood? 

In a detailed technical seminar, AbbVie scientists Shitalben Patel and Dr. Jami Erickson presented their internal development of two novel CETSA® applications: Flow Cytometry CETSA and Quantitative Whole-Blood CETSA. 

Note: These methods were developed by AbbVie for their internal pipeline using a Pelago license. While not currently offered as off-the-shelf Pelago services, they represent the cutting edge of what is possible with CETSA® technology. 

Flow Cytometry CETSA: Single-Cell Resolution 

Dr. Erickson presented a high-throughput (384-well) screen targeting C-RAF. Unlike traditional bulk-lysis methods, this flow-based approach maintains cell integrity. 

• The Scientific Edge: By using gating strategies, AbbVie can analyze target engagement in specific cell subpopulations. 

• Sensitivity: In a screen of ~2,500 compounds, the Flow CETSA format identified known inhibitors, whereas comparative assays missed several, proving superior sensitivity for the C-RAF target. 

Whole-Blood CETSA: Bypassing PBMC Isolation 

Shitalben Patel addressed the “translational gap” with a protocol designed for clinical utility. Using RIPK1 as a proof-of-concept target, the team developed homogeneous assays using MSD (Meso Scale Discovery) and AlphaLISA platforms. 

• Sample Economy: The assay provides robust EC₅₀ data using <100 µL of whole blood, eliminating the variability and labour associated with PBMC isolation. 

• Clinical Robustness: Crucially, the team validated that the “CETSA effect” (thermal shift) remains stable after freeze-thaw cycles. This allows for asynchronous sample collection at clinical sites and batched analysis at a central facility. 

• Correlation: The results showed a strong positive correlation with traditional Receptor Occupancy assays, validating the biological relevance of the data. 

AbbVie’s work illustrates the immense flexibility of the CETSA® principle. By adapting the readout methods (Flow and MSD/Alpha), they have created a seamless translational workflow. They can now validate hits with single-cell precision in discovery, and confirm engagement in complex blood matrices in the clinic. 

Watch the Technical Presentation 

Access the full webinar to view the melting curves, gating strategies, and correlation data presented by the AbbVie team. 

You are welcome to contact Pelago in Japan to discuss how CETSA can empower your program. stefan.sandstrom@pelagobio.com