Microscale Thermophoresis System

Monolith X
Monolith X is a system based on microscale thermophoresis (MST) technology to measure binding affinity between biomolecules in solution, without the need for immobilization. It is a tool for characterizing small molecule-protein, protein-protein, and protein-nucleic acids interactions, as well as for the study of protein aggregation concentration determination.

  • Spectral Shift Technology to quantify molecular interactions between a target and a ligand. When the target is labeled with a fluorophore, it gives a particular emission spectrum. If a ligand binds, the fluorophore’s chemical environment is changed, causing a blue-shift or redshift or broadening of the emission peak. Fluorescence is recorded and a 670 nm/650 nm ratio is calculated and plotted against the ligand concentration. This ratiometric measurement is used to derive the affinity constant (Kd) which is automatically determined at the end of each run without additional and lengthy data analysis.
  • MST technology to understand aggregation. It measures variations in fluorescence intensity caused by ligand binding while a very precise and brief laser-induced temperature change is applied. The change in fluorescence is then plotted against the concentration of the binding partner to obtain the dissociation constant (Kd).
  • Combination of MST, sensitive to the presence of aggregates in samples, and Spectral Shift in the same instrument, to provide two modalities that complement each other to get further insights about molecular interactions.
  • Samples that can be analysed: biological macromolecules (proteins, nucleic acids, lipids) or low molecular weight molecules (e.g. organic, inorganic molecules, sugars).
  • Measurements in aqueous solutions (buffer solutions).
  • Ultra-low sample consumption.
  • Time to get Kd: 10 min (Spectral Shift + MST); 90 seconds (Spectral Shift).
  • Dynamic range: nM to mM.
  • Detected molecule range: 101-107 Daltons.
  • Temperature control: 20-40 °C +/- 0.5 °C (actively controlled).
  • Fluorescent channels: 1 (RED).

📍The system is located at Elettra Sincrotrone Trieste laboratories in Area Science Park.

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