The objective of this study was to determine differences in composition and functionality for oat protein concentrates. This was achieved by comparing preparations extracted at different pH levels using either a twin screw press or alkaline extraction followed by isoelectric precipitation. Extraction by twin screw technology provided a higher protein yield compared to isoelectric precipitation. Analysis by SDS-PAGE revealed similar protein composition regardless of the extraction method or extraction pH except for extraction at pH 6. Extraction at a higher pH led to a higher protein purity and yield, while extraction closer to the isoelectric point of the protein resulted in less pure concentrates. All extracts were freeze-dried, and their composition and functionality were compared. Thermal analysis of the protein concentrates extracted by twin screw pressing showed a higher denaturation enthalpy compared to protein isolates prepared by isoelectric precipitation, suggesting that the native structure of the proteins is preserved to a higher extent. All protein fractions exhibited the typical U-shaped solubility profile, except for the concentrate extracted at pH 6, which showed an increase in solubility with increasing pH. Oat protein concentrates prepared by twin screw pressing had a higher affinity for binding to oil than to water, and this was also higher than for isoelectric precipitated protein extracts. Overall results demonstrate a potential for twin screw technology as a viable alternative to conventional wet extraction methods for producing less refined fractions from oats with tunable functional properties.