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Biofouling of membrane surfaces by the attachment of microorganisms is one of the major obstacles for ensuring the effectiveness of membrane separation processes. This work presents the construction of a zwitterionic PVDF membrane surface with improved resistance to biofouling. An amphiphilic copolymer of poly(vinylidene fluoride)-graft-poly(N,N-dimethylamino-2-ethylmethacrylate) (PVDF-g-PDMAEMA) was first synthesized via radical graft copolymerization and then the flat membrane was cast with immersed phase inversion. The PDMAEMA side chains tended to aggregate on the membrane surface, pore surface and internal pore channel surface, and were converted with 1,3-propane sultone (1,3-PS) to yield a zwitterionic membrane surface. A higher conversion of PDMAEMA chains and distribution of zwitterions were obtained using a longer treatment time. A biofouling assay indicated that incorporation of zwitterions suppressed the adsorption of extracellar polymer substances and the adhesion of Escherichia coli bacterial cells to the membrane surface, endowing the membrane with a high flux recovery and biofouling resistance in the filtration process.