Herein, polyethersulfone (PES) and sulfonated polysulfone (SPSf) blend membranes were prepared with addition of sulfonated polyethersulfone (SPES) as a hydrophilic polymer and adipic acid as a porogen via non-solvent induced phase separation method for effective fractionation of dyes based on the influence of steric hindrance and charge effect. Raman spectroscopy and molecular dynamic simulation modeling confirmed that hydrogen bonds between PES, SPSf, SPES, and adipic acid were crucial to membrane formation and spatial arrangement. Further addition of hydrophilic SPES resulted in a membrane with reduced pore size and molecular weight cut-off as well as amplified negative charge and pure water permeance. During separation, the blend membranes exhibited higher rejection rates for nine types of small molecular weight (269.3–800 Da) dyes than for neutral polyethylene glycol molecules (200–1000 Da). This was attributed to the size effect and the synergistic effect between steric hindrance and charge repulsion. Notably, the synergistic impact decreased with dye molecular weight, while greater membrane negative charge enhanced small molecular dye rejection. Ideal operational stability and anti-fouling performance were best observed in M2 (PES/SPSf/SPES, 3.1 wt %). Summarily, this study demonstrates that SPES with −SO₃⁻ functional groups can be applied to control the microstructure and separation of membranes.