The TNKS2 series was selected for additional testing because: it is part of a newer extended benchmark set that has been less studied than the FEP+ set (which includes TYK2); it involves fewer ring changes than TYK2; it contains multiple R-group sites at different sections of the ligand scaffold. Note that six +1 net charge ligands (8a-f) were excluded from this series as charge perturbations were considered out of scope for this work, the series included n=21 ligands. For TNKS2 only the RBFENN and LOMAP-Score network edges were simulated in quintuplicates (see figure and S15 for RBFE networks). For this series, a 'dynamic' representation is used to investigate statistical performance when adding replicates (figure ). Similar performance was observed between the two networks, with a similar MUE of 0.9 kcal•mol -1 when including all replicates (figure ). This similarity is conserved when including fewer replicates, with little statistical difference between the two approaches.The same holds true for Pearson R and Kendall τ (figures 10A and B, resp.). The dynamic representation of statistical performances across replicates for TNKS2 highlights the importance of of assessing protocol repeatability. . It appears that none of the statistical metrics have fully reached a plateau after 5 repeats, suggesting that the RBFE protocol could benefit from an even larger number of replicates or other optimisations. Although in this analysis no reference can be made to an optimal network chosen according to |∆∆G offset | values as in section 3.3.4, the RBFENN (n=28) and LOMAP-Score (n=27) networks can be directly compared. With an overlap of 27%, the overlap is considerably lower than with TYK2. The eight shared edges are predominantly single-atom perturbations.