MicroScale Thermophoresis: Interaction analysis and beyond

2014/03/20 by Moran Jerabek-Willemsen, Moran Jerabek‐Willemsen, Timon André +6 · 714 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · Engineering · #Chemical physics #Chemistry #Computational chemistry #Field-Flow Fractionation Techniques #Heat shock proteins research #Microscale chemistry #Microscale thermophoresis #Molecular dynamics #Molecule #Nanoparticle #Nanotechnology #Organic chemistry #Small molecule #Thermophoresis #thermodynamics and calorimetric analyses

paper · pdf · doi:10.1016/j.molstruc.2014.03.009

published in Journal of Molecular Structure 1077, 101-113 (Elsevier BV)

openalex publication_date 2014/03/20 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01

Abstract

MicroScale Thermophoresis (MST) is a powerful technique to quantify biomolecular interactions. It is based on thermophoresis, the directed movement of molecules in a temperature gradient, which strongly depends on a variety of molecular properties such as size, charge, hydration shell or conformation. Thus, this technique is highly sensitive to virtually any change in molecular properties, allowing for a precise quantification of molecular events independent of the size or nature of the investigated specimen. During a MST experiment, a temperature gradient is induced by an infrared laser. The directed movement of molecules through the temperature gradient is detected and quantified using either covalently attached or intrinsic fluorophores. By combining the precision of fluorescence detection with the variability and sensitivity of thermophoresis, MST provides a flexible, robust and fast way to dissect molecular interactions. In this review, we present recent progress and developments in MST technology and focus on MST applications beyond standard biomolecular interaction studies. By using different model systems, we introduce alternative MST applications – such as determination of binding stoichiometries and binding modes, analysis of protein unfolding, thermodynamics and enzyme kinetics. In addition, wedemonstrate the capability of MST to quantify high-affinity interactions with dissociation constants (Kds) in the low picomolar (pM) range as well as protein–protein interactions in pure mammalian cell lysates.

Citations

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