In developed countries, the highest number of amputees are elderly with transtibial amputation. Walking on inclined surfaces is difficult for amputees due to loss of muscle volume and strength thereby transtibial amputees (TA) rely on the intact limb to maintain stability. The aim of this study was to use the concatenated non-negative matrix factorization (CNMF) technique to calculate muscle synergy components and compare the difference in muscle synergies and their associated activation profiles in the healthy and amputee groups during ramp ascending (RA) activity. Healthy subjects' dominant leg and amputee's intact leg (IL) were considered for recording surface electromyography (sEMG). The muscle synergies comparison showed a reasonable correlation between the healthy and amputee groups. This suggests the central nervous system (CNS) activates the same group of muscles synergistically. However, the activation coefficient profile (C) results indicated statistically significant difference (p <; 0.05) in some parts of the gait cycle (GC) in healthy and amputee groups. The difference exhibited in activation profiles of amputee's IL could be due to the instability of the prosthetic leg during the GC which resulted in alteration of the IL muscles activations. This information will be useful in rehabilitation and in the future development of prosthetic devices by using the IL muscles information to control the prostheses.