MicroScale Thermophoresis (MST) is a technique that researchers use to analyze interactions between biomolecules. It uses the change in fluorescence between two molecules to assess biomolecule interactions. MicroScale Thermophoresis is a powerful procedure based on the movement of molecules in a temperature gradient.
MST enables researchers to determine binding affinity without needing to purify the target protein. This cuts out several difficult and time-consuming steps. The MST method uses an overexpression of the green fluorescent protein (GFP), as well as cell breakdown and the detection of microscale thermophoresis signals in the presence of ligand concentrations.
Determining binding affinity is essential in many different life science studies – especially during the creation of new drugs. Using the straightforward microscale thermophoresis protocol, rather than outdated research techniques, can save time and resources by allowing the study of binding affinity without first dealing with protein purification. It is a new way to analyze proteins and understand binding affinity. As a researcher, measuring binding affinity might be useful for a variety of applications.
Thanks to MST technologies, researchers can now easily measure interactions between different molecules, using very little sample. You can measure binding affinity in minutes and without wasting any of the protein sample. One way in which you can use microscale thermophoresis is to study protein-peptide interactions.
Studying protein-peptide interactions is important in drug design and biochemistry. With MST, it is now faster and easier than ever to determine the most important parameters of an interaction between biomolecules – including thermodynamics and dissociation constant. You do not need to worry about sample immobilization or having enough sample for consumption with this technique.
MST also suitable for analyzing protein folding and unfolding processes. The folding properties of different proteins are available for study using this new method. The study of the chemical denaturation (unfolding) of proteins is important to understand their folding properties and overall stability. These studies are particularly important if the researcher needs to design or optimize therapeutic biomolecules, or gain a deeper understanding of diseases that relate to protein-misfolding.
MST excels at analyzing protein unfolding. It rivals traditional approaches due to faster speeds and lower protein sample consumption, while providing the same information. It is possible to use microscale thermophoresis to monitor and characterize the simple to complex chemical denaturation of proteins. This technique can also detect unfolding intermediates without interfering with the stability of the protein.
Drug discovery processes are increasingly considering the rational optimization of molecular interactions. This marks another application for MST, specifically in single dose and affinity screening. While it’s not a direct measurement, thermodynamics can be considered. It can be derived from the affinity data and can then be used to acquire additional mechanistic information. Thermodynamic parameters are used to reveal the molecular mechanism of an interaction, and that information can be used for the rational design of interactions. This leads to the discovery of new drugs.
The next time you need to study a protein sample, consider using MST to obtain accurate results with efficiency.