The application of sum frequency generation (SFG) vibrational spectroscopy to investigate protein behavior through the study of deuterated proteins has been demonstrated for the first time, showing the overall importance and promise of this combination of techniques. Using deuterated proteins and SFG, we report the feasibility of molecular level studies on the responses of polymer surfaces to protein adsorption, the dynamic process of protein adsorption, and protein mixture adsorption. We also demonstrate that the combination of selective protein isotope labeling and the SFG technique can be expanded in the future to elucidate more detailed structures of adsorbed proteins.