NanoDSF

nanoDSF is a modified Differential Scanning Fluorimetry method to determine protein stability employing intrinsic tryptophan fluorescence.^[1][2] nanoDSF was developed by NanoTemper Technologies GmbH, a German high tech company with headquarters in Munich.

Protein stability is typically addressed by thermal or chemical unfolding experiments.^[3] In thermal unfolding experiments, a linear temperature ramp is applied to unfold proteins, whereas chemical unfolding experiments use chemical denaturants in increasing concentrations. The thermal stability of a protein is typically described by the 'melting temperature' or 'Tm', at which 50% of the protein population is unfolded, corresponding to the mid-point of the transition from folded to unfolded.

In contrast to conventional DSF methods, nanoDSF uses tryptophan fluorescence to monitor protein unfolding. Both the fluorescence intensity and the fluorescence maximum strongly depends on the close surroundings of the tryptophan.^[4] Therefore, the ratio of the fluorescence intensities at 350 nm and 330 nm is suitable to detect any changes in protein structure, for example due to protein unfolding.

Thermal unfolding curves and unfolding transition midpoints of two monoclonal antibodies. (A) Thermal unfolding curves of two monoclonal antibodies in the presence of 25mM Na-Citrate at different pH vlaues. Insets show the pH-dependence of the first unfolding midpoint (Tm1). (B) Dependence of Tm1 and Tm2 on the pH of the buffer of all tested conditions.

Its applications include antibody engineering, membrane protein research, quality control and formulation development.^[5]

References

External links

• NanoTemper Technologies