Recent advances in molecular simulation methods and greater availability of computing resources allow much more comprehensive exploration of biomolecular systems at the atomistic level than was previously possible. A number of previous studies have shown molecular dynamics (MD) simulations to be a powerful tool for understanding the properties and dynamics of insulin. Mark et al. and Zoete et al. found that the insulin monomer is more flexible than the dimer. Zoete et al. additionally confirmed the high flexibility in the B-chain C-terminus of insulin, which was consistent with the experimental data. Later, Yang et al. used simulated annealing to probe how PEGylation enhanced the stability and potency of insulin, discovering that an optimal chain length existed for PEGglyated insulin pharmaceuticals. More recently, with steered molecular dynamics and replica-exchange umbrella sampling , Antoszewski et al. revealed different pathways of insulin dimer dissociation and characterized relevant energetic and structural features of the process. Busto-Moner et al. presented a pipeline integrating parallel tempering , the integrated variational approach for conformational dynamics (IVAC) , Markov state model (MSM) , and Perron cluster analysis (PCCA) for sampling the wild-type insulin ensemble and they concluded three important elements of disorder common in the insulin ensemble at a low pH value.