In case functional assays are available TriCEPS coupled ligands can also be tested to see if a similar output can be achieved with a TriCEPS coupled ligand compared to a ligand that is not coupled to TriCEPS.
As first step, the TriCEPS v.2.0 molecule is coupled to the ligand of interest (peptide, protein, Antibody, ADC or other primary amine containing molecules) and to the positive control ligand (e.g. transferrin) and negative control ligand (e.g. glycine). This coupling reaction is tested with Dot blot to assess if the coupling worked.
Then, the TriCEPS coupled ligands are added to the non-oxidized cells to assess whether the ligand binds to the unknown targets at the cell surface and TriCEPS does not interfere with the ligand receptor interaction.
Flow Cytometry tests with TriCEPS coupled ligand on oxidized cells can be further performed to test cell viability and ligand target binding as it will occur in the main LRC-TriCEPS experiment.
After successful completion of all pretests, the main experiment can be planned and carried out as described in the LRC-TriCEPS user manual choosing one of the service options provided by Dualsystems Biotech AG.