(2) Apply filtering, for instance through multiple sequence alignment, or tools like our partner tool Key Interaction Networks (KIN), to identify evolutionary conserved positions or interactions. Modifications at these positions will likely be highly detrimental to activity. (3) Once a manageable number of hotspots have been filtered, these can be subject to randomization using computational tools such as FuncLib, to predict promising variants, which can finally (4) be subjected to further experimental design and/or testing. In this way, steps 1 and 2 could, in principle, replace the NMR component of the design process.