We performed MicroScale Thermophoresis (MST) measurements to determine the dissociation constants of protease and substrate. We benefited from the low sample consumption, the small volumes and the possibility to choose the buffer freely especially to work at high salt concentrations. Furthermore MST allowed us to optimize assays in a straight forward manner and to carry out fast measurements resulting in highly reproducible data. The dissociation constants of USPs and various substrates measured by MST are in very good agreement with previously published affinities using alternative approaches like ITC, FA (fluorescence anisotropy) and SPR.
PD Dr. Günter Fritz
University of Freiburg