For instance, the MUE for the entire dataset using a single replica ranges from 1.19 ± 0.05 to 1.24 ± 0.05 kcal/mol, whereas it is 1.04 ± 0.04 kcal/mol for TIES. The corresponding numbers for the subset |∆∆G exp | < 1 kcal/mol are 1.00 ± 0.05 to 1.06 ± 0.05 kcal/mol versus 0.92 ± 0.04 kcal/mol. Similar trends are observed for r p values. It is worth highlighting that the number of ligand pairs with |∆∆∆G| < 1 kcal/mol (that is, an absolute difference between experimental and predicted relative free energies of less than 1 kcal/mol) using ensemble simulations is higher than those using a single replica (296 versus 262-276). Thus, ensemble simulations yield binding free energy predictions for 20 to 34 more ligands with accuracy better than the 1 kcal/mol mark compared to one-off simulations.