A minimal thermodynamic theory for re-entrant liquid-liquid phase separation regulated by small molecules

Small molecules regulate biomolecular condensates in a biphasic manner, promoting liquid–liquid phase separation (LLPS) at low concentrations while suppressing it at higher concentrations. Despite increasing experimental evidence for such re-entrant behavior, a unified physical description remains lacking. Here, we identify a minimal thermodynamic mechanism for re-entrant LLPS by coupling Cahn–Hilliard dynamics to a concentration-dependent Flory interaction parameter containing competing LLPS-promoting and inhibitory contributions. The resulting model reproduces experimentally observed nonmonotonic condensate formation in Tau–tannic acid and TDP-43–bis-ANS systems, including the concentration-dependent emergence and dissolution of protein-rich domains. Spinodal analysis reveals finite concentration windows for phase instability and demonstrates that re-entrant mixing is encoded directly in the free-energy landscape. The framework further captures morphology transitions and diffusive coarsening within the phase-separated regime. These results establish a general mesoscale description of chemically regulated condensates and provide design principles for controlling phase separation through small-molecule modulators.