The effect of the addition of triblock copolymer polyethylene glycol (PEG)–polypropylene glycol (PPG)–polyethylene glycol (PEG) (Pluronic F127) and polyethylene glycol (PEG-4000, M_n = 4000 g mol⁻¹) to the polysulfone (PSF) casting solution on the membrane structure and performance was studied. The phase state, viscosity and turbidity of PSF solutions in N,N-dimethylacetamide (DMAc) with the addition of block copolymer Pluronic F127 were investigated. It was found that 18–22 wt.% PSF solutions in DMAc with Pluronic F127 content ≥ 5 wt.% feature a lower critical solution temperature (LCST). Membrane structure was investigated using scanning electron and atomic force microcopies. It was revealed that average pore size and pore amount on the surface of the membrane selective layer increase and pore size distribution becomes wider with an increase in Pluronic F127 content in the casting solution. It was found that the average surface roughness parameters of the membrane selective layer for PSF/Pluronic F127 membranes significantly exceed those for PSF/PEG-4000 membranes. It was shown that the increase in the membrane flux and the decrease in polyvinylpyrrolidone (PVP K-30, M_n = 40,000 g mol⁻¹) rejection are a result of the addition of both Pluronic F127 and PEG-4000 into the casting solution. It was revealed that PSF/Pluronic F127 membranes are characterized by higher pressure resistance in ultrafiltration process, a lower total flux decrease during ultrafiltration of bovine serum albumin solutions. The antifouling performance of PSF/Pluronic F127 membranes was found to exceed significantly the antifouling performance of PSF/PEG-4000 membranes.