iONCscan Services for Co-Stimulatory and Inhibitory Checkpoint Receptors

Accelerate Cancer Immunotherapy Drug Development by Outsourcing Your Screening and Potency Needs

Regulation of immune responses is tightly controlled through a balance of co-stimulatory and inhibitory checkpoint receptors, often exploited by many cancers. Therefore, therapeutics that block inhibitory receptors or activate immune-stimulatory checkpoint receptors are powerful agents to restore anti-tumor immune responses, such as anti-PD-1 therapy.  However, developing drugs targeting these checkpoint proteins is quite challenging as cell-based assays used to screen for functional drugs are often difficult to create, involve the use of primary cells, and have long, complicated protocols.  To solve this bottleneck, Eurofins DiscoverX’s iONCscan Services offer an expanding menu of cell-based assays that can enable screening & potency determination for immunotherapy drugs.

Advantages of iONCscan Services for Checkpoint Receptors
  • Potency determination – Rank order potency of your molecules
  • Fast turnaround time – Obtain results in 30 days without the need to develop and validate assays in-house
  • Multiple applications – Can be used for biologic and small molecule drug development

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Product ▲Cell LineLigandPart No.
PathHunter® Jurkat CD200R Signaling Cell LineJurkat PathHunter® Jurkat CD200R Signaling Cell Line
PathHunter® Jurkat CD28 Signaling Cell Line JurkatAnti-CD28 Antibody[CD28.2]PathHunter® Jurkat CD28 Signaling Cell Line
PathHunter® Jurkat CTLA4 Signaling Cell LineJurkat PathHunter® Jurkat CTLA4 Signaling Cell Line
PathHunter® Jurkat ICOS Signaling Cell LineJurkatAnti-ICOS antibodyPathHunter® Jurkat ICOS Signaling Cell Line
PathHunter® Jurkat PD-1 (SHP1) Signaling Cell LineJurkatAnti-PD1 Antibody [NAT105]PathHunter® Jurkat PD-1 (SHP1) Signaling Cell Line
PathHunter® Jurkat PD-1 (SHP2) Signaling Cell LineJurkat PathHunter® Jurkat PD-1 (SHP2) Signaling Cell Line
PathHunter® Jurkat SIRPa Signaling Cell LineJurkat PathHunter® Jurkat SIRPa Signaling Cell Line
PathHunter® U2OS CD137 Signaling Cell LineU2OS4-1BB-LPathHunter® U2OS CD137 Signaling Cell Line
PathHunter® U2OS FcgRIa Clustering Cell LineU2OS PathHunter® U2OS FcgRIa Clustering Cell Line
PathHunter® U2OS FcgRIIa Clustering Cell LineU2OS PathHunter® U2OS FcgRIIa Clustering Cell Line
PathHunter® U2OS FcgRIIb Clustering Cell LineU2OS PathHunter® U2OS FcgRIIb Clustering Cell Line
PathHunter® U2OS OX40 Signaling Cell LineU2OSOX40LPathHunter® U2OS OX40 Signaling Cell Line
PathHunter® U2OS PD-1/PD-1 Dimerization Cell LineU2OSAnti-PD1 Antibody [NAT105] PathHunter® U2OS PD-1/PD-1 Dimerization Cell Line
PathHunter® U2OS TIM3/TIM3 Dimerization Cell LineU2OSAnti-TIM3 Antibody [344801]PathHunter® U2OS TIM3/TIM3 Dimerization Cell Line
PathHunter® U2OS VISTA/VISTA Dimerization Cell LineU2OSAnti-VISTA Antibody [730804]PathHunter® U2OS VISTA/VISTA Dimerization Cell Line
  • Measure potency of your molecules against custom-selected checkpoint family targets or every available checkpoint assay target

  • Screen collections or a small number of molecules against one or more custom selected checkpoint assays

Reproducibility with PathHunter iONCscan Services

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’ tab above.

Better Sensitivity, Quicker Results than Reporter Gene Assays

PathHunter PD-1 Assay vs. Competitor
Assay PathHunter Assay Competitor Assay
IC50 ng/mL 54 927
Assay Time < 5 hours 22 hours
Signal:Background Ratio 11.1 7.8

Using the same anti-PD-1 antibody (BioLegend Cat. No. 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 better assay window, and returned results in one-quarter the total assay time. 

Screen and Identify Both Small Molecule and Biologic Drugs

PD-1 is known to be phosphorylated by an as yet unidentified Src family kinase, which results in recruitment of the SHP-1 protein to the phosphorylated tail of the 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.

Example Principle of PD-1 Signaling Assay

Download scientific poster on the PD-1 assay to view additional data

Ligand Dimerization Principle

The two target receptors are tagged with ProLink™ (PK) or Enzyme Acceptor (EA). Upon ligand-induced activation, the receptors dimerize forcing the two β-gal components to complement and create an active enzyme. Active β-gal generates a chemiluminescent signal in the presence of substrate.