"Label-Free & Dye-free Characterization of Protein Stability and Aggregation using nanoDSF" 

Wyatt Strutz
Senior Application Scientist
NanoTemper Technologies Inc.

We will introduce the Prometheus NT.48 instrument and its novel nanoDSF technology, which allows for parallel high-precision characterization of stability and aggregation parameters of biologicals. nanoDSF is an advanced Differential Scanning Fluorimetry technology. It detects smallest changes in the fluorescence of tryptophan present in virtually all proteins.

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 an ultra-high resolution unfolding curves which allow for detection of even minute unfolding signals. Furthermore, since no secondary reporter fluorophores are required as in conventional DSF, protein solutions can be analyzed independent of buffer compositions, and over a concentration range of 200 mg/ml down to 5 μg/ml. 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.