Notably, both the proteolytic stability and dimerization propensity are related to transformation processes that require overcoming free energy barriers. The proteolytic stability can be directly associated with the free energy barrier of the formation of the conformational ensemble corresponding to the transition state in the proteolytic degradation by 𝛼-chymotrypsin. Similarly, dimerization propensity is strongly correlated with the free energy barrier to dimerization. While these free energy differences could serve as more direct measures for the two biophysical properties of interest, calculations of such free energy differences are far from trivial. The reason lies in the fact that the timescale of the binding/unbinding events between an insulin monomer and an 𝛼-chymotrypsin or between two insulin monomers are prohibitively long. This necessitates the use of advanced sampling techniques to accelerate the sampling of the configurational space of the system of interest, such as umbrella sampling or alchemical transformations . However, such free energy methods are usually much more complicated and computationally expensive than standard MD simulations of the same length. The increase in the system size of insulin/protease complex or insulin dimer calculations adopting either method could easily increase the computational cost significantly beyond the scope of what is possible to screen large numbers of insulin modifications. Advanced configurational sampling approaches can often be significantly difficult to set up and interpret, thus making easy screening impossible.