The earthquake in northeastern Japan that occurred on March 11, 2011 brought about the nuclear accident, resulting in the detection of radioactive cesium in soil and water over a wide region around Fukusihma. In this study, with the aim of the establishment of an effective method for removing cesium from water contaminated with cesium, the functionalized membrane with large cesium adsorption capacity per unit mass was prepared by combining nanofibers having a large specific surface area with iron ferrocyanide having a high selectivity for cesium adsorption. The nanofiber membrane made of polyacrylonitrile (PAN) was used as a base material of the functionalized membrane. Nanofiber membranes were immersed in the dispersions of iron ferrocyanide with various concentrations and pH values. After taking it out, it was dried at various temperatures and then non-immobilized iron ferrocyanide was completely removed through cleaning. As a result of the evaluation of completed affinity membranes, the amount of iron ferrocyanide immobilized by the nanofiber membrane increased significantly with the increase in the iron ferrocyanide concentration but subsequently showed a tendency to decrease rapidly, resulting in a distinct maximum at the iron ferrocyanide concentration of 3 wt%. And, the supported amount of iron ferrocyanide to the nanofiber membrane increased as pH became lower. Moreover, it was found that as high temperature as possible without exceeding the glass transition temperature of PAN was optimal as a drying temperature of prepared affinity membrane.