Figure summarizes the main findings: in absolute terms, MD-based non-equilibrium free energy calculations perform comparably to conventional in silico predictors, with an overall adjusted predictive AUE of 0.68±0.05 pK taken as an average over each residue class (compared to 0.74±0.07 pK and 0.70±0.06 pK for the consensus of the Poisson-Boltzmann (PB) methods and empirical (EM) PropKa method, respectively) (Figure ). Regarding the individual residue classes computed using the MD approach, for the adjusted estimate, AUEs were: 0.77±0.09 pK (aspartate), 0.69±0.09 pK (glutamate) and 0.52± 0.04 pK (lysine) (Figure ).