To enable such deconvolution several computational algorithms have evolved that map the binding site using a variety of molecular probes at the protein surface. Commonly used programs such as MCSS or GRID have now been superseded by FTMap (E-FTMap) PLImap and Hotspots API . These methods broadly differ in their general utility of our approach to direct fragment-based design studies, across sixteen different protein systems. We also assess the quality of the predictions and introduce the application of dynamic hotspots prediction named Dy-FMOPhore for flexible protein systems.