In order to develop novel membranes for vanadium redox flow battery (VRB) with low self-discharge rate and low cost, sulfonated poly(fluorenyl ether ketone) (SPFEK) was synthesized directly via aromatic nucleophilic polycondensation of bisphenol fluorene with 60% sulfonated difluorobenzophenone and 40% difluorobenzophenone. The SPFEK membrane shows the lower permeability of vanadium ions. The open circuit voltage evaluation demonstrates that the SPFEK membrane is superior to Nafion 117 membrane in self-discharge test. Both energy efficiencies (EE) and power densities of the VRB single cell based on the SPFEK membrane are higher than those of the VRB with Nafion 117 membrane at the same current densities. The highest coulombic efficiency (CE) of VRB with SPFEK membrane is 80.3% while the highest CE of the VRB with Nafion 117 membrane is 77.0%. The SPFEK membrane shows the comparative stability to Nafion 117 membrane in VO2 + electrolyte. The experimental results suggest that SPFEK membrane is a promising ion exchange membrane for VRB.