Interest in alternative plant-based protein sources is continuously growing. Sugar beet leaves have the potential to satisfy that demand due to their high protein content. They are considered as agricultural waste and utilizing them as protein sources can bring them back to the food chain. In this study, isoelectric-point-precipitation, heat-coagulation, ammonium-sulfate precipitation, high-pressure-assisted isoelectric-point precipitation, and high-pressure-assisted heat coagulation methods were used to extract proteins from sugar beet leaves. A mass spectrometry-based proteomic approach was used for comprehensive protein characterization. The analyses yielded 817 proteins, the most comprehensive protein profile on sugar beet leaves to date. Although the total protein contents were comparable, there was a significant difference between the methods for low-abundance proteins. High-pressure-assisted methods showed elevated levels of proteins predominantly located in the chloroplast. Here we showed for the first time that the extraction/precipitation methods may result in different protein profiles that potentially affect the physical and nutritional properties of functional products.