One assay – three formats 

Pelago Biosciences offers 3 assay formats for our customers. Pelago has developed the different formats of CETSA into tailored services for different applications to meet customer needs. In all cases an initial feasibility assessment is made by our CETSA specialists and customers will be advised on suitability before an order is necessary.


CETSA Classics

The CETSA classics format allows for rapid assay validation and generation of EC50’s to confirm ligand potency and ranking. It utilizes a Western Blot-based detection for target quantification.  Typically used for confirmation of target engagement of 1-15 compounds in for example human cell lines, primary cells, tissues, xenografts, bacteria, insects and crops. The assay development phase is typically 3-4 weeks with an additional 2-3 weeks for subsequent EC50 screening.


Our high-throughput plate-based format is ideal for library screening, hit confirmation and lead optimisation. The plate-based proximity detection is automation ready. Typically, the HT format has an assay development phase of 8-10 weeks. Completion of subsequent EC50 screens will depend on volume. Alternatively, the CETSA-HT assay can also be exported to the customer after the assay development phase.


CETSA MS, often referred to as TPP (Thermal Proteome Profiling) and more recently as PISA (Protein Integral Solubility Alteration), allows detection of target engagement of thousands of proteins in a single experiment, building an in-depth drug profile by identifying compound binding events and off-target interactions. Once the experimental parameters have been specified, the turnaround time is 6-12 weeks depending on format. This is the recommended assay format for biomarker investigation, toxicological profiling and target de-convolution.

Proteins of interest uncovered by CETSA MS are amenable to both Classics and HT methods. This allows for the generation of specific assays suitable for low or high throughput once targets have been identified.


Real world EC50 values for your compound in cells 

CETSA Classics and CETSA HT employ antibody-based detection, the assays are suitable both in lysate and intact cells, and can be applied to a wide variety of target classes.­

Once an assay has been developed for a target, the translation into alternative matrices is often straight forward. This allows compounds identified early in the project to be investigated in other matrices, for example animal tissues and patient samples to confirm preliminary results. 


Un-Biased Proteomic Profiling

CETSA MS, also known as thermal proteome profiling, is a mass spectrometry based derivation of CETSA. It can analyse the systemic effect of a compound on up to 6,000 key proteins. Measuring not just the direct binding of compound to proteins but also the subsequent downstream consequences of the initial target engagement. This unbiased proteome wide profiling makes it ideal for target de-convolution, toxicity & biomarker studies.

Proteins of interest uncovered by CETSA MS are amenable to Classics and HT methods and therefore specific assays suitable for low-high throughput can be developed once targets have been identified.

Contact us to discuss your project needs

For a video introduction of CETSA Classics, CETSA HT and CETSA MS, watch the video below.