Overview
Interaction analysis by the SUPR-DSF offers high-speed and label-free measurements across the analyte binding concentration range. The measurements are free in solution and are not influenced by surface effects, mass transport or refractive index buffer issues. The measurements confirm binding is specific by detecting induced stability changes in the protein-ligand complex. This is depicted from the shift in the emitted fluorescence as it melts, measured as a function of ligand concentration, from which the KD can be calculated. This method uses commercially available, low cost, low volume 384-well microplates, and is capable of screening thousands of samples for stability in a day to confirm binding. Binding induced stability changes are readily apparent even for very weak binding partners (e.g., fragment libraries), confirming the binding is specific and of structural/functional importance. The SUPR-DSF can be integrated into existing robotic solutions, making very large-scale screens easy and rapid to complete. The below Investigation applies the SUPR-DSF platform to the study of binding effects of a ligand with carbonic anhydrase.
