Advantages Of The Checkpoint Assays
- No primary cells – Get biologically relevant responses without the use of difficult and costly primary cells
- Simple Protocol, Fast Results – Increase efficiency with an add and read protocol and results in less than 5 hours
- Highly Sensitive Response – 15X more sensitive than other assays
- Multiple Applications – Monitor checkpoint signaling for biologic and small molecule drug development
Subtract primary cells. Add reproducibility and ease of use.
The PathHunter PD-1 Checkpoint Assay was performed with Pembrolizumab (anti-PD-1 antibody) and PD-L1 ligand presenting cell line. Data show consistent EC50’s and S/B on three different runs of the assay within the same plate with less than 4% relative standard deviation (%RSD). For more details on how the assay works, please visit the ‘Assay Principle & Data’ tab above.
Complete full assay in less than 5 hours with simple, add and read protocol.
15X more sensitive than other reporter gene assays
Using the same anti-PD-1 antibody (BioLegend Catalog #329912), we were able to compare the assay performance of our PathHunter PD-1 signaling assay to a commercially available reporter gene assay, and observed that the PathHunter PD-1 assay demonstrated more than 15-fold better sensitivity than the reporter gene assay, with a slightly better assay window as well.
Screen and identify both small molecules and biologic drugs
PD-1 is known to be phosphorylated with an as yet unidentified src family kinase, which causes the recruitment of the SHP-1 protein to the phosphorylated PD-1 receptor. Testing of the PathHunter PD-1 assay with small molecule inhibitors that are known to inhibit several known src family kinases has shown inhibition of the SHP-1 recruitment to PD-1, demonstrating that the PathHunter PD-1 assay can be used to identify novel small molecule inhibitors of this exciting immunotherapy target.