Microscale Thermophoresis (MST)
Microscale thermophoresis (MST) is a technique for biophysical analysis of biomolecular interactions. MST is based on the detection of temperature induced target fluorescence changes as a function of the concentration of non-fluorescent ligands. MST can be used with almost all kinds of target molecules, if they are fluorescent or can be labeled with fluorescent dyes. The diversity of ligands is greater. The fluorescent target molecule keeps a constant concentration, and the non-fluorescent ligand molecule titrates in a large concentration range.
MST is carried out in a thin capillary in free solution, thus providing near natural conditions (no immobilization in any buffer, even in complex biological liquids) and maintenance free instruments. In MST experiment, infrared laser can cause micro temperature gradient, and detect tric and thermophoresis. Tric depends on the microenvironment of the fluorophore and usually changes during binding events. The movement of molecules in a temperature gradient, thermophoresis, depends on three parameters that usually vary with the interaction. Therefore, the dose-response curve can be obtained by plotting the whole MST signal relative to the ligand concentration, from which the binding affinity can be deduced.
Applications
- Single dose screen: MST will tell you whether the target and ligand usually interact through a single dose experiment.
- Steady state affinity: MST will tell you the exact and true steady-state affinity of the interaction (dissociation constant Kd).
- Protein aggregation: MST will tell you whether the protein sample is aggregated or whether the aggregation is induced by ligand binding.
- Ligand competition: MST will tell you the competitive effect of multiple adhesives targeted.
Figure 1. Multiple applications of MS (Jerabek-Willemsen, M.; et al. 2014)
Services
As a protein research expert, Profacgen offers various services using MicroScale Thermophoresis technology. We have introduced high-end NanoTemper equipment to meet the requirements of customers for rapid and high-throughput screening. A constant concentration of the fluorescent probed molecule and a variable concentration of any titrant are required for each experiment. Our MST platform is based on a 96-capillary-based system that requires only a small amount of sample per capillary (just nM concentrations and µL volumes), which saves protein usage. On this platform, you can not only determine affinity, but also assess other physical parameters such as stoichiometry, aggregation, enthalpy (van't Hoff plot), slow enzyme kinetics and oligomerization, among others. Our MST services can also be combined according to your project needs to help solve all problems in protein analysis, drug discovery, and antibody development.
Advantages
- Low sample consumption: only 4µl is required per sample
- Free choice of assay buffers: biological liquids can also be used
- Short analysis times: fast and high throughput
- Real-time quality controls: online polymerization, precipitation and sticking control
- Readily allows competitive binding studies to be performed, and orthosteric and allosteric binders to be discovered
- No immobilization required: measurement is done in solution
- Wide temperature range: analysis can be performed from 20°C to 45°C
- Wide concentration range: Affinities can be analyzed in the pM-mM range
- Wide molecular size range: from 100 Da to 1 MDa
As a protein research expert, Profacgen has accumulated a lot of successful experience in protein interaction analysis, and has assisted many customers around the world to complete projects with high efficiency and high quality. Our highly competitive prices and unprecedented expertise have earned us widespread acclaim. Contact us to find out how Profacgen could be of assistance.
Reference
- Jerabek-Willemsen, M.; et al. MicroScale Thermophoresis: Interaction analysis and beyond. Journal of Molecular Structure. 2014.