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Isothermal titration calorimetry and thermal shift assay in drug design

20 June 2011  •  Author(s): Asta Zubrienė, Egidijus Kazlauskas, Lina Baranauskienė, Vytautas Petrauskas and Daumantas Matulis, Department of Biothermodynamics and Drug Design, Vilnius University Institute of Biotechnology

Isothermal titration calorimetry (ITC) is a method of choice in the pharmaceutical industry for determination of equilibrium binding enthalpy, entropy, and the Gibbs free energy. The method is very powerful for determination of intrinsic binding parameters that could be used in structure-energetics correlations. Here we discuss how to overcome several limitations of ITC. First, it is easy to complement ITC results with thermal shift assay (TSA) in order to avoid the narrow window of ITC Kd measurements. Second, several examples are provided on how to determine intrinsic enthalpy of binding and Kd. Third, ITC and TSA Kds are compared with enzymatic inhibition methods.

Isothermal titration calorimetry is a wellestablished method for determining the association constant and other thermodynamic parameters of intermolecular interactions in aqueous solutions. The technique has been widely used to study interactions between very different molecular species such as proteins, DNA, RNA, lipids, drug leads, metal ions and many other chemical substances as previously reviewed1,2.

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