𝛼-chymotrypsin is a common digestive protease secreted by the pancreas which performs proteolysis in the duodenum . Experimental work used 𝛼-chymotrypsin to assess the proteolytic stability by measuring the half-life of different insulin glycoforms. In proteolytic degradation of insulin by 𝛼-chymotrypsin, residues including TyrA14, TyrA19, TyrB16, PheB25, and TyrB26 serve as the cleavage sites, whose scissile bonds are on the carboxyl side . Previous research into 𝛼-chymotrypsin indicated that residues PheB25 and TyrB26 were the cleavage sites considered to be the most susceptible to proteolytic stability. We therefore hypothesized that the exposure of the scissile bonds of these two residues to the solvent was positively correlated with proteolytic susceptibility.