How does the inherent asymmetry of the locomotor system in people with lower-limb amputation affect the ankle-foot roll-over shape of prosthetic walking? In a single-case design, we evaluated the walking patterns of six people with lower-limb amputation (3 transtibial and 3 transfemoral) and three matched nondisabled controls. We analyzed the walking patterns in terms of roll-over characteristics and spatial and temporal factors. We determined the level of asymmetry by roll-over shape comparison (root-mean-square distance) as well as differences in radius of curvature. In addition, we calculated ratios to determine spatial and temporal asymmetries and described different aspects of asymmetry of roll-over shapes. All participants showed some level of asymmetry in roll-over shape, even the nondisabled controls. Furthermore, we found good intralimb reproducibility for the group as a whole. With respect to spatial and temporal factors, the participants with transtibial amputation had a quite symmetrical gait pattern, while the gait in the participants with transfemoral amputation was more asymmetrical. The individual ankle-foot roll-over shapes provide additional insight into the marked individual adjustments occurring during the stance phase of the nondisabled limb. The two methods we present are suitable for determining asymmetry of roll-over shapes; both methods should be used complementarily.