Potato protein is recognized as one of the most valuable nonanimal proteins due to the high content of essential amino acids. So far, it has not been used in human nutrition on a large scale due to technological limitations regarding its acquisition. In this study, the protein fraction of potato juice was concentrated with the use of membrane separation. The obtained potato juice protein concentrate (PJPC) was characterized in terms of nutritional value and biological activity, and the amino acid composition, mineral content, and antioxidant properties were determined. Moreover, in vitro cytotoxic activity against cancer cells of the gastrointestinal tract was investigated. The results of the present study indicate that PJPC is an excellent source of lysine and threonine, while leucine is its limiting amino acid, with an amino acid score (AAS) of 65%. Moreover, PJPC contains substantial amounts of Fe, Mn, K, and Cu. As demonstrated experimentally, PJPC is also characterized by higher antioxidant potential than potato itself. Biological activity, however, is not limited to antioxidant activity alone. Cytotoxicity studies using a gastric cancer cell line (Hs 746T), a colon cancer cell line (HT-29), and human colon normal cells (CCD 841 CoN) proved that PJPC is characterized by selective activity against cancer cells. It can thus be concluded that the developed method of producing protein concentrate from potato juice affords a product with moderate nutritional value and interesting biological activity.