The fluorescence of tryptophans in a protein is strongly
dependent on its close surroundings. By following changes in fluorescence,
chemical and thermal stability can be assessed in a truly label-free fashion.
The dual-UV technology by NanoTemper allows for rapid fluorescence detection,
providing an unmatched scanning speed and data point density. This yields
ultra-high resolution unfolding curves which allow for detection of even minute
unfolding signals. Furthermore, since no secondary reporter fluorophores are
required, protein solutions can be analyzed independent of buffer compositions,
and over a concentration range of 150 mg/ml down to 5 µg/ml. In addition,
information on protein aggregation can be recorded in parallel, providing
insight into colloidal stability of the sample. Therefore, nanoDSF is the
method of choice for easy, rapid and accurate analysis of protein folding and
stability, with applications in membrane protein research, protein engineering,
formulation development and quality control.
The presentation will cover biophysical concepts of the
technique showing benefits of the nanoDSF technology platform, and will be
followed by specific examples of nanoDSF applications towards various
experimental systems.