The effect of high hydrostatic pressure (HHP) application on whey protein concentrate was evaluated both before (pre-treatment - PT) and during (hydrolysis assisted - HA) hydrolysis processes. A factorial design 22 with 3 central points was used with pressure (100, 250, 400 MPa) and time (5, 20 and 35 minutes) as independent variables. The hydrolysis was evaluated and monitored by soluble protein, aromatic amino acid contents and RP-HPLC. ABTS and ORAC tests were used to evaluate the in vitro antioxidant capacity. The reduction of soluble protein content was approximately 20% for conventional hydrolysis and for all PT treatments up to 4 h of reaction, while HHP assisted hydrolysis at 100 MPa showed a 35% protein reduction after 35 minutes of reaction. In addition, pressurization favored peptic hydrolysis of β-lactoglobulin by up to 98% and also improved the in vitro antioxidant capacity of the hydrolysates, which increased from 34.25 to 60.89 μmoles TE g-1 of protein in the best treatment. The results suggest that the use of HHP assisted hydrolysis favored the peptic hydrolysis, with a reduction in hydrolysis time and increased antioxidant activity.