Technical Abstract: Some milk-associated proteins are known to be nutritionally valuable and form bioactive peptides that exhibit activity against hypertension and oxidative stress. Consumption of cheeses, such as the popular Hispanic-style cheese Queso Fresco (QF), may be a vehicle for delivery of these milk-associated peptides. This paper describes the effects of high-pressure processing (HPP) on the antioxidative activity (ORAC-FL value) of water-soluble proteins extracted from QF samples. QFs were manufactured according to a commercial-make procedure using pasteurized, homogenized milk, without added starter cultures. The cheese was cut into 45x45x150mm3 blocks, double packaged in vacuum bags, and received the following HPP treatments: 200, 400, or 600 MPa for either 0, 5, 10, or 20 min, with warming to an internal temperature of either 22 or 40 deg C prior to HPP treatment. Results show that the core temperature of the cheese during HPP directly affects the ORAC-FL value. The activities of the lower temperature cheeses are time and pressure, and have a median ORAC-FL value of 27 trolox equivalents (TE). The higher temperature cheeses have higher ORAC-FL values ranging from 21.5 to 96.0 TE; the highest activity corresponded to the cheese held at 400 MPa for the longest time under pressure (20 min). The 600 MPa cheeses increase in activity with increasing time under pressure, but are less active than the control cheese. These results indicate that processing temperature and pressure are important factors in the antioxidative activity of these QF samples and further understanding of the roles of these variables may lead to the manufacture of healthier and more nutritious cheeses and dairy products.