NanoTemper MST

Microscale Thermophoresis for Quantitative Analysis of Molecular Interactions

Microscale Thermophoresis (MST) is a solution-based biophysical technique for the quantitative analysis of molecular interactions. At the Fisher Drug Discovery Resource Center, NanoTemper MST instrumentation is used to measure interactions between proteins, small molecules, and other binding partners using very small sample volumes.

MST detects changes in the movement of molecules through a microscopic temperature gradient. Binding of a ligand can alter this thermophoretic behavior, allowing interactions to be measured directly in solution.

Applications in Drug Discovery

MST is particularly useful for characterizing and quantifying molecular interactions, including:

  • Determination of binding affinities (KD)
  • Characterization of protein–small molecule interactions
  • Confirmation and follow-up of screening hits
  • Comparison of compound analogs and structure–activity relationships
  • Analysis of interactions involving proteins, nucleic acids, peptides, and other biomolecules

Because measurements are performed in solution and require very small sample volumes, MST can be especially valuable when purified target protein is available only in limited quantities.

Advantages

MST combines quantitative binding measurements with low sample consumption and relatively rapid assay development. Measurements are performed under solution conditions, which can allow interactions to be studied in an environment closer to the native state than some immobilization-based techniques.

Key advantages include:

  • Very low sample consumption
  • Quantitative determination of binding affinity
  • Measurements performed in solution
  • Minimal sample preparation compared with many other biophysical methods
  • Suitable for a broad range of molecular interactions
  • Useful for hit confirmation and compound characterization

MST is therefore a versatile tool for moving from initial screening results to quantitative characterization of compound–target interactions.