Techniques and skills
Thermophoresis, or Soret effect, refers to a phenomenon based on the movement of particles or molecules subjected to a temperature gradient. The thermophoresis of a compound varies depending on its size, charge, or solvation level according to the terms of Soret’s equation. During a specific interaction between two molecules, at least one of these properties is affected (size, charge, solvation) and consequently modifies the thermophoresis curve.
Based on the principle of thermophoresis, micro-scale Thermophorése or MicroScaleThermophoresis (MST) allows the demonstration of molecular interactions between biomolecules of different physico-chemical natures, purified or not.
Detection of variations in the mobility of a fluorescent molecule (naturally or by fluorescent labeling) subjected to a micro-gradient of temperature established in a capillary, in the presence of increasing concentrations of a specific interactant.
The exploitation of the thermophoretic properties of a compound for molecular interaction studies constitutes an innovative approach.
On the PCBI platform, we have the Monotih NT.115 from Nanotemper to perform these analyses.
The nanoDSF is the method of choice for fast and accurate measurement of protein stability. It detects changes in the fluorescence of tryptophan, a hydrophobic amino acid present in proteins. This fluorescence is highly dependent on the environment close to this amino acid. In a protein with correct conformation, it is located in the hydrophobic core of the protein and protected from the aqueous environment of the solvent. During the unfolding, tryptophan becomes exposed, which alters its fluorescence. The unfolding of proteins can thus be recorded in a real approach without prior labeling.
On the PCBI platform, we have the TYCHO TM NT.6 from Nanotemper to perform these analyses.
Proteins are essential elements. Recombinant proteins are proteins (secreted, nuclear, membrane) produced in the laboratory by a bacterium or by eukaryotic cells (yeast, insect cells or mammals), whose genetic material has been modified.
After purification, these proteins can be used to understand complex cellular mechanisms or in therapies.
The PCBI platform has a cell bank (CHO, HEK293...) as well as access to an L2-type laboratory in order to carry out cell culture and recombinant protein production. The recombinant proteins produced can be used in MST interaction studies (with a ligand, with another protein or with a small molecule).
