MagHelix™ Microscale Thermophoresis (MST)

Microscale thermophoresis (MST) has become a common technique to detect specific target-probe interactions, and it measures the differences in the movement rate through a microscopic temperature gradient caused when complexes are formed. MST can be applied to characterize any type of biomolecular interactions, for example, protein-DNA, protein-RNA, protein-protein, antigen-antibody interactions, as well as the binding of a ligand to ternary complexes.

With the advantages of high sensitivity, low sample consumption, and short experiment time, MST technology is suitable for high-throughput screening (HTS) and fragment-based screening (FBS) in the early stage of drug discovery. Creative Biostructure has been one of the first enterprises to master the latest techniques with forward-looking insights into the development of structure-guided drug discovery. We have established many reliable approaches to meet your drug discovery program requirements. We can provide MST, a relatively new methodology for quantifying biomolecular interactions to assist screen and characterize hit compounds.

Brief Introduction to Microscale Thermophoresis (MST)

MST is a non-immobilized technique for quantitative analysis of biomolecular interactions in solution. The technique is based on thermophoresis and fluorescence detection. The instrument of MST utilizes an infrared laser for local heating to cause molecular directional movement, and then analyze the molecular distribution ratio in the temperature gradient field by fluorescence (fluorescence labeling or intrinsic fluorescence). The MST technique can detect ligand binding-caused changes in thermophoretic mobility, which rely on size, charge, hydration shell, and conformations. Kd values can be estimated using these changes in thermophoretic mobility.

MagHelix™ Microscale Thermophoresis (MST)Figure 1. The schematic of MST technology. (Alexander C G.; et al. 2014)

In short, MST has many advantages in the classification and characterization of hits. For example, 1) simple and easy to operate, does not require immobilization and purification of target proteins (fluorescent fusion proteins); 2) Low sample consumption and experimental operation cost; 3) Real-time acquisition of affinity data (such as dissociation constant); 4) This measurement can be performed in a variety of different solutions, including liposomes or detergents needed to study membrane proteins. However, MST depends on the fluorescence of either the compound or the targe, and because the fluorophore itself may affect certain aspects of binding and therefore requires validations.

Advantages of our MagHelix™ Microscale Thermophoresis (MST) services:

  • We have many years of experience in using MST technology to study biomolecular interactions, and our MST services have been proven to be reliable by customers from biotech, pharmaceutical, and biopharmaceutical industries.
  • Determination of binding stoichiometries and the number of binding sites.
  • Determination of thermodynamic parameters of interactions, such as ΔG (Gibbs free energy), ΔH (enthalpy), and ΔS (entropy).
  • We can apply MST technology to perform high-throughput screening of compounds/fragments and fragment-based screening.
  • By combining with other biophysical technologies, the limitations of MST are compensated to make the results more reliable.

As an advanced contract service provider in the drug discovery phase, Creative Biostructure can simultaneously apply a variety of established approaches and biophysical techniques to identify, validate, classify and characterize the hit compounds according to the goals and requirements of the project. If your project has such needs, please feel free to contact us, and our scientists will provide professional consultation for you.

References

  1. Alexander C G.; et al. Novel microscale approaches for easy, rapid determination of protein stability in academic and commercial settings. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics. 2014, 1844(12): 2241-2250.
  2. Bartoschik T.; et al. Microscale thermophoresis in drug discovery. Applied Biophysics for Drug Discovery. 2017: 73-99.
  3. Rainard J M.; et al. Using microscale thermophoresis to characterize hits from high-throughput screening: a European lead factory perspective. SLAS DISCOVERY: Advancing Life Sciences R&D. 2018, 23(3): 225-241.
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