MST is the biophysical technology that facilitates the analysis of interactions between biomolecules.
Based on the detection of a temperature-induced change in fluorescence of a target as a function of the concentration of a non-fluorescent ligand.
The observed change in fluorescence is based on two distinct effects: principally via a temperature related intensity change (TRIC) of a fluorescent probe, which is affected by binding or conformational change events; and thermophoresis… the directed movement of particles in a microscopic temperature gradient.
Any change of the chemical microenvironment of a fluorescent probe, result in a relative change of the fluorescence detected when a temperature gradient is applied and can be used to determine binding affinities.
The extent of this change in fluorescence intensity is affected by the chemical environment of the fluorescent probe… …this can be altered in binding events due to proximity of ligands.
Concentration dependence of this quenching effect is directly dependent upon the binding affinity of the ligand.