nanoDSF – Technology
nanoDSF – a tool to study melting and aggregation onset temperatures
The nanoDSF is a differential scanning fluorimetry method able to analyze the conformational stability and colloidal stability (aggregation behavior) of proteins under different thermal and chemical conditions.
The conformational stability of a protein is described by the unfolding transition midpoint Tm (°C), which is the point at which half of the protein is unfolded. The truly labelfree nanoDSF monitors the intrinsic fluorescence of tryptophanes in proteins, which rely on the close surrounding of the amino acid and changes upon thermal unfolding (read more about the theoretical background here).
In addition, the nanoDSF can determine the colloidal stability (aggregation behavior) of proteins by backreflection optics. Therefore the loss of light, which passes the protein sample, is reflected by the capillary tray and passes the sample again is used as a measure for the aggregation onset temperature of proteins. (read more about the theoretical background here)
Figure: technical setup nanoDSF:
Up to 48 capillaries are filled with up to 10 µl of protein sample and simultaneously scanned at 330 / 350 nm wavelength for melting temperatures and with backreflection optics for aggregation onset temperatures while being heated up from 15°C to 95°C. Within 75 min melting temperatures and aggregation onset temperatures of 48 conditions can be studied without the use of a dye and with free choice of buffer and detergent. Melting temperatures of proteins with a concentration between 5 micro g / ml up to 250 mg / ml can be analyzed using the nanoDSF. To obtain high quality aggregation onset temperatures, the protein is commonly used in concentrations above 1 mg / ml.
nanoDSF is a rapid and accurate method to study the conformational and colloidal stablity of proteins and is thus perfectly suited for protein engineering, formulation development and screening approaches
Read more about the advantages of the nanoDSF.
For more information on the nanoDSF, please also watch this video about the nanoDSF.
We offer to characterize your protein of interest using the nanoDSF
If you are interested in nanoDSF services please read more here