In this work， neohesperidin (NH) and whey protein (WP) complex (NH-WP) was prepared to explore the possibility of enhancing the antioxidant activity of whey protein (WP). The non-covalent interaction mechanism of NH with WP was studied in terms of fluorescence spectroscopy， synchronous fluorescence spectroscopy， thermodynamic analysis and molecular docking simulation. The antioxidant activity of NH-WP was also analyzed by a series of common used methods. The results showed that the endogenous fluorescence of WP could be statically quenched by NH， which changed the microenvironment of aromatic amino acids in WP. Non-covalent interactions between NH and WP lead to the formation of NH-WP complexes， characterized by a negative ΔG， positive ΔS， and positive ΔH， suggesting an endothermic and spontaneous process primarily driven by hydrophobic forces. There was only one binding site between them and the complex was formed by combining them at a ratio of 1∶1. In addition， the antioxidant activity of NH-WP complex was lower than that of free NH at the same concentration， but higher than that of the pure protein， what's more， the antioxidant activity of WP exhibited dose-dependent effect with the increase of NH addition， suggesting that the introducing of NH could effectively improve the antioxidant activity of WP. This research could enrich the theoretical basis of the interaction between WP and polyphenols， and provide good reference basis for improving functional properties of proteins as well as widening the application of NH and WP in the field of food and medicine.