To validate our algorithm, we compared the residues detected and categorized during the apo-structure scan and the holo-complex analysis across sixteen different systems, classifying them into hotspots and non-hotspots. By quantifing the recall of hotspots identified in the development of binders (holo-complex) for each target, if they were correctly identified as hotspots using static FMOPhore mode in the apo-structure scenario. The recall between the holo-complex structures for each system and its equivalent in the apo-structure is calculated using the following equation ( ):