Electrospun nanofiber membranes (ENMs) have demonstrated promising applications for water purification primarily due to high water flux and low degree of fouling. However, the equivalent/apparent pore sizes of as-electrospun ENMs are in microns/submicrons; therefore, the ENMs can only be directly utilized for microfiltration applications. To make regenerated cellulose (RC) ENMs for ultrafiltration applications, atom transfer radical polymerization (ATRP) was studied to graft polymer chains onto the surface of RC nanofibers; specifically, monomers of 2-hydroxyethyl methacrylate (HEMA) and sodium acrylate (AAS) were selected for surface-grafting water-insoluble and water-soluble polymer chains onto RC nanofibers, respectively. With prolonging of the ATRP reaction time, the resulting surface-modified RC ENMs had reduced pore sizes. The water-insoluble poly(HEMA) chains coated the surface of RC nanofibers to make the fibers thicker, thus decreasing the membrane pore size and reducing permeability. On the other hand, the water-soluble poly(AAS) chains did not coat the surface of RC nanofibers; instead, they partially filled the pores to form gel-like structures, which served to decrease the effective pore size, while still providing elevated permeability. The surface-modified RC ENMs were subsequently explored for ultrafiltration of ∼40 nm nanoparticles and ∼10 nm bovine serum albumin (BSA) molecules from water. The results indicated that the HEMA-modified RC membranes could reject/remove more than 95% of the nanoparticles while they could not reject any BSA molecules; in comparison, the AAS-modified RC membranes had complete rejection of the nanoparticles and could even reject ∼58% of the BSA molecules.