This work studied the interactions between trehalose/chitooligosaccharide (COS) and myofibrillar protein (MP), and the effect of such interactions on the quality of meat after freezing was also evaluated. Fourier transform infrared spectroscopy showed that both trehalose and COS could enhance the content of hydrogen bonds of MP. Zeta potential measurement displayed trehalose/COS reduced the absolute value of the surface potential of MP. The results of Raman spectroscopy suggested that the hydrophobic residues of MP were more exposed after treatment with trehalose/COS. Thus, trehalose and COS could both interact with MP through non-covalent bonds. Subsequently, the evaluation of the effect of trehalose and COS on the physicochemical properties of frozen meat was conducted. Results showed that both trehalose and COS significantly reduced thawing loss of frozen meat, and sensory evaluation showed that trehalose had a better performance from the perspective of smell, texture, and overall consumer acceptance. In conclusion, trehalose/COS interacting with MP can reduce meat thawing loss, which might provide technical guidance in the quality control of frozen meat.