Isothermal ligand binding analysis

Sigurskjold, B.W. (2000) Exact analysis of competition ligand binding by displacement isothermal titration calorimetry, Anal.Biochem. 277, 260-266 ... 

Freyer, M. W., Lewis, E. A. (2008). Isothermal titration calorimetry experimental design, data analysis, and probing macromolecule/ligand binding and kinetic inteac-tions. Methods Cell Biol., 84, 79-113. 

Chaires JB, Analysis and interpretation of ligand-DNA binding isotherms. Methods Fnzymol. 2001 340 3-22. 

From Equation (17) an analysis can be made if the quantity of ML can be determined as a function of ligand added at each aliquot. However considerable errors may arise as this quantity must be assayed, after each aliquot of ligand is added. However non-invasive techniques such as isothermal microcalorimetry calorimetry can be used to directly determine the quantity of ML without disturbing the system. For a calorimetric analysis an estimation of the enthalpy change for the macromolecule-ligand interaction must be made. An assumption can be made that at the start of the study, the initial aliquot of ligand added to the system, if sufficiently small, will completely bind to the macromolecule (as initially the macromolecule will be in excess). The enthalpy change associated with this interaction can then be used to calculate the A// for the interaction (Equation (3)). 

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