Simultaneous determination of thermodynamic and kinetic data by isothermal titration calorimetry

• Published in: Biochimica et biophysica acta. General subjects Vol. 1865; no. 2; p. 129772
• Main Authors: Glöckner, Steffen, Klebe, Gerhard
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2021
• Subjects: 1:1 protein-ligand complexes; Animals; calorimetry; Calorimetry - methods; Cattle; Drug Discovery; Drug optimization; drugs; Humans; Kinetic binding data; Kinetics; kinITC; Ligands; Measurement protocol; Protein Binding; Proteins - metabolism; Surface Plasmon Resonance; Thermodynamic binding data; Thermodynamics; titration; 1:1 protein-ligand complexes; kinITC; Measurement protocol; Drug optimization; Thermodynamic binding data; Kinetic binding data
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2020.129772

Thermodynamic and binding kinetic data increasingly support and guide the drug optimization process. 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. 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. 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’. 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. [Display omitted] •Thermodynamic and binding kinetic data for 1:1 protein-ligand complexes recorded in one experiment under same conditions•No labeling or immobilization required•Material- and time-efficient hybrid protocol that accounts for protein samples of reduced activity•Required material for measurements is the same as for the usual thermodynamic analysis