In this study, we examined the preparation and hydrolysis property of immobilized urease membrane to decompose harmful urea in the body and remove ammonia which was produced by its decomposition. Urease immobilized membrane was prepared by introducing anion-exchange group DEA into porous hollow-fiber membrane by radiation graft polymerization method, and immobilization of urease. When urease was immobilized at membrane introduced with anion-exchange group, the more increasing grafting rate, the more increasing immobilization amount. The result originates from the fact that a greater amount of protein was immobilized by forming a multilayer on the longer grafted chain. Meanwhile, the addition of the cross-linker was possible not only to suppress separation phenomenon produced during a washing process of immobilized urease membrane but also to enable the recycling of membrane. Urease Immobilized membrane with no separation phenomenon was prepared by cross-linking reaction for 5 h, and the hydrolysis rate of prepared urease immobilized membrane was over 98% and 50%, respectively, in 1 mol and 4 mol urea solutions.