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Updated: Nov 30, 2025

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions
Published on: April 4, 2014
Simultaneous determination of thermodynamic and kinetic data by isothermal titration calorimetry
Steffen Glöckner1, Gerhard Klebe1
1Philipps-University Marburg, Institute for Pharmaceutical Chemistry, Marbacher Weg 6, 35037 Marburg, Germany.
Background:
Thermodynamic and binding kinetic data increasingly support and guide the drug optimization process.
Methods:
Because ITC thermograms contain binding thermodynamic and kinetic information, an efficient protocol for the simultaneous extraction of thermodynamic and kinetic data for 1:1 protein ligand reactions from AFFINImeter kinITC in one single experiment are presented.
Results:
The effort to apply this protocol requires the same time as for the standard protocol but increases the precision of both thermodynamic and kinetic data.
Conclusions:
The protocol enables reliable extraction of both thermodynamic and kinetic data for 1:1 protein-ligand binding reactions with improved precision compared to the 'standard protocol'.
General Significance:
Thermodynamic and kinetic data are recorded under exactly the same conditions in solution without any labeling or immobilization from a protein sample that is not 100% active and would otherwise render the extraction of kinetic parameters impossible.
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