MicroScale Thermophoresis – Comparison

Comparing the MicroScale Thermophoresis to other biophysical methods

Important methods used to investigate molecular interactions include Isothermal Titration Calorimetry (ITC), Surface Plasmon Resonance (SPR), and Fluorescence Correlation Spectroscopy (FCS). Comparing datasets obtained by these technologies with MicroScale Thermophoresis data reveals high similarity. This indicates that the output data of these technologies are absolutely comparable. However, MicroScale Themophoresis assays provide valuable advantages as shown in the summary below.

 

FACTS & FEATURES: MicroScale Thermophoresis compared to other methods
MethodMicroScale Thermophoresis (MST)
Affinity Range (Kd)
  • pM to mM
Advantages
  • measurements in solution
  • free choice of reaction conditions
  • low material consumption
  • temperature controlled
  • fast measurements
  • interaction between small and big molecules detectable
  • fluorescence and labe-free assays
  • multiplex assay
Draw Backs
  • Kon and Koff rates are not accesible
MethodSurface Plasmon Resonance (SPR)
Affinity Range (Kd)
  • sub nM to low mM
Advantages
  • high sensitivity
  • method of choice measuring Kon and Koff rates
Draw Backs
  • lab-intense establishment of new assays
  • covalent coupling might interfere binding event
  • artefacts stemming from mass transport limitations close to an interface
MethodIsothermal Titration Calorimetry (ITC)
Affinity Range (Kd)
  • nM to sub mM
Advantages
  • method of choice to measure thermodynamic parameters of an interaction
Draw Backs
  • high material consumption
  • high preparation time
  • buffer limitations
  • low to medium throughput
  • interactions with no or only a small change in enthalpy not measurable
Method Fluorescence Correlation Spectroscopy (FCS)
Affinity Range (Kd)
  • low nM to mM
Advantages
  • high sensitivity
  • in vivo measurements possible
Draw Backs
  • low throughput
  • high preparation time
  • large changes in molecular masses required for detection

For detailed informations, download the 2bind – MicroScale Thermophoresis booklet