On carefully observing Figure (top), it can be noted that when considering all long Figure : ADRP-tofacitinib complex with splitting protocol: (A) Distribution of ligandresidue contact frequencies for each set of sub-replicas, (B) Volume occupancy maps of the ligand around protein rendered at an isovalue of fractional occupancy 0.03. For each set of subreplicas, the wireframe isosurface represents the area of the simulation box where the ligand is likely to be found with 97% probability. trajectories in aggregate, we are able to recover all ligand-protein interactions which are identified across the splitting protocol. To see this more clearly, we plotted the ligand-residue contact frequency distributions for aggregated sampling time from both the splitting protocol (9 µs in total) as well as unbiased sampling (100 µs in total) for ADRP system in Figure (bottom). Here we see that the concatenated trajectory reproduces all modalities which occur across the splitting protocol, albeit with different statistical weights. Incidentally, if our aim is to simply explore all possible binding sites and dominant poses within those sites, the unbiased long timescale protocol is far less efficient than the splitting protocol which achieves this aim in an aggregate of 1 day and 7 hours of wall clock time rather than 43 days 2 hours of wall clock time required with the former. However, care must be taken when thermodynamic quantities need to be evaluated/predicted using the splitting protocol as the biased sampling leads to biased weights of the microstates sampled that may affect the averages obtained.