9
How it works
In contrast to conventional DSF methods in which
fluorescence labeling is required, nanoDSF measures
intrinsic tryptophan or tyrosin fluorescence to monitor
protein unfolding. Both the fluorescence intensity and
the fluorescence maximum strongly depends on the close
surroundings of the tryptophan. 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. It can be used to characterize
thermal stability or unfolding, chemical denaturation, and
relative aggregation.
Strengths
With this technique, protein stability can be monitored
under native conditions. Also, no sample prep is needed
and little sample is consumed. Another beneficial
feature of the technology is the robust data density or
the number of readings collected during each data point
that translates into higher quality and more reliable
information collected during each experiment compared
to other methodologies.
Weaknesses
There are trade-offs when considering nanoDSF vs other
methodologies. There are methods that provide higher
throughput (DSF/Thermofluor) but require labeling of
samples and don't provide highly quantitative results.
There are systems available that can perform multiple
analysis on protein stability, however, the precision of each
method may be compromised vs. nanoDSF. Researchers
need to consider the quality and amount of data generated
and required when evaluating the weaknesses of nanoDSF.
Conclusion
NanoDSF is a label-free approach which uses rapid sample
scanning and intrinsic fluorescence detection to collect
highly precise and accurate thermal unfolding profiles in a
high-throughput manner.
Nano differential scanning fluorimetry (nanoDSF) is a modified version of DSF that measures protein stability using
intrinsic tryptophan or tyrosin fluorescence.
nanoDSF
