Microscale thermophoresis for the analysis of biomolecular interactions

The present proposal aims to equip several research groups from three
research institutes (LDRI, IREC, DDUV) and one research centre
(NAMEDIC) of two universities of the Wallonie-Bruxelles Federation (UCL
and UNamur) with a microscale thermophoresis instrument (MST). Briefly,
MST allows for quantitative analysis of protein interactions in free solution
and with low sample consumption. The technique is based on
thermophoresis, the directed motion of molecules in temperature gradients.
Thermophoresis is highly sensitive to all types of binding-induced changes
of molecular properties, be it in size, charge, hydration shell or
conformation. In an all-optical approach, an infrared laser is used for local
heating, and molecule mobility in the temperature gradient is analysed via
fluorescence. In standard MST one binding partner is fluorescently labeled.
However, MST can also be performed label-free by exploiting intrinsic
protein UV-fluorescence.
It should be noted that currently none of these laboratories or institutes have
access to this technology. However, the measurement of biomolecular
interactions and the subsequent derivatisation of dissociation constant (Kd)
in a very efficient and rapid way being increasingly important in modern
drug discovery, we are convinced that the acquisition of an MST instrument
would greatly facilitate and accelerate our research.