Nano differential scanning fluorimetry

NanoDSF is a type of differential scanning fluorimetry (DSF) method used to determine conformational protein stability by employing intrinsic tryptophan or tyrosine fluorescence, as opposed to the use of extrinsic fluorogenic dyes that are typically monitored via a qPCR instrument.

In contrast to conventional DSF methods, nanoDSF uses tryptophan or tyrosine fluorescence to monitor protein unfolding.

The latter BCM method takes advantage of the entire UV-fluorescence spectrum, thus allowing for flexibility when auto-fluorescent small molecules are present.

[8] Currently there are at least four instruments on the market that can measure fluorescence wavelength shifts in a high-throughput manner while heating the samples through a defined temperature ramp.

[13] NanoDSF was used to compare the thermal stability of a matched set of anti-CD20 antibodies representing a range of variants.

Thermal unfolding curves and unfolding transition midpoints of two monoclonal antibodies. (A) Thermal unfolding curves of two monoclonal antibodies in the presence of 25 mM Na-Citrate at different pH values. 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.