Resistant Potato Starch (RPS) is a fundamental prebiotic used in dairy products. The study aims to evaluate the role of RPS as a fat substitute in the production of Labneh cheese. RPS1, RPS2, and RPS3 labneh treatments were prepared using 0.5, 1, and 1.5% of RPS, compared to modified buffalo milk full-fat control labneh (FBC) 4% fat, and low-fat control labneh (LBC) 2% fat. The physicochemical, microbiological, and sensory characteristics of labneh were investigated after 29 days of storage. The results demonstrated that RPS2 and RPS3 had remarkable significant differences (P > 0.0001) in physicochemical and sensory properties. As a consequence, we recommend that RPS may be used in the manufacturing of functional labneh.