Estimation of drug–protein binding parameters on assuming the validity of thermodynamic equilibrium

This contribution focuses the reader’s attention on the pitfalls usually emerging during the phase of evaluation of experimental data of drug–protein binding studies. To overcome the occurrence of problem(s) apparently defying solution, the concept of “affinity spectra” is recommended to be implemented for data evaluation. A (general) “binding study protocol” is also suggested, which can prevent the formation of inadequate conclusions and the generation of unrealistic drug–protein binding parameters.