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The use of biophysical methods for characterizing ligand binding to macromolecules of biological interest is presented in terms of three questions: Who? Where? Why? The “who” question addresses the problem of identification of the ligands binding to a given target, the “where” question addresses the problem of determining the site of binding, and the “why” questions try to establish the physical (especially thermodynamic) basis of binding. This chapter establishes this framework, which will be followed in the rest of the book.

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