MicroScale Thermophoresis detects interactions between any kind of biomolecules thus providing a large application range, from ions and small molecules to high molecular weight and multi-protein complexes.
Thermophoresis, the movement of molecules in temperature gradients, is not only dependent on the size, but also on the charge and the hydration shell of the molecule of interest. Therefore, binding events can be detected even without an increase in size or mass upon complex formation.
Extensive research conducted at the Biophysics Department of the Ludwig-Maximilians-University Munich (LMU) (1) identified the solvation entropy and the hydration shell of molecules as the driving force. Any change of the hydration shell of biomolecules due to changes in their primary, secondary, tertiary and/or quaternary structure affects the thermophoretic movement and is used to determine binding affinities with high accuracy. The dependency on the energetic of the hydration shell allows for sensitivity unmatched by any other technology.
Since MST is performed free in solution without any surface immobilization, also bulky or sensitive molecule assemblies such as liposomes, nanodiscs or membrane proteins can be investigated.
Employing MST, the affinity between any kind of biomolecules but also other biophysical parameters such as stoichiometry, binding energetics, unfolding, can be determined.
|(1)||Seidel, S. A. I., P. M. Dijkman, W. A. Lea, et al.Microscale thermophoresis quantifies biomolecular interactions under previously challenging conditions. Methods59: 301-315; (2013). Goto reference|