Protein stereostructure analysis in mixtures still remains challenging, especially large-scale analysis such as in proteomics. With the capability of measuring the hydrodynamic radius of ions in the liquid phase, mobility capillary electrophoresis (MCE) has been applied to study the structure of peptides. In this study, MCE was extended for protein mixture separation and their corresponding hydrodynamic radius analyses. After ellipsoid approximation, the results obtained by MCE experiments were then used as a restraint in molecular dynamics simulations to predict the most probable structure of each protein. Besides a three-protein mixture, a mixture of disulfide bond reduced insulin was also studied by this MCE-restrained modeling method. The results obtained by this method agree with literature studies, and mass spectrometry experiments were also carried out to confirm our findings.