This study was aimed to compare the variability of inter-joint coordination in the lower-extremities during gait between active individuals with transtibial amputation (TTAs) and healthy individuals (HIs). Fifteen active male TTAs (age: 40.6 ± 16.24 years, height: 1.74 ± 0.09 m, and mass: 71.2 ± 8.87 kg) and HIs (age: 37.25 ± 13.11 years, height: 1.75 ± 0.06 m, and mass: 74 ± 8.75 kg) without gait disabilities voluntarily participated in the study. Participants walked along a level walkway covered with Vicon motion capture system, and their lower-extremity kinematics data were recorded during gait. The spatiotemporal gait parameters, lower-extremity joint range of motion (ROM), and their coordination and variability were calculated and averaged to report a single value for each parameter based on biomechanical symmetry assumption in the lower limbs of HIs. Additionally, these parameters were separately calculated and reported for the intact limb (IL) and the prosthesis limb (PL) in TTAs individuals. Finally, a comparison was made between the averaged values in HIs and those in the IL and PL of TTAs subjects. The results showed that the IL had a significantly lower stride length than that of the PL and averaged value in HIs, and the IL had a significantly lower knee ROM and greater stance-phase duration than that of HIs. Moreover, TTAs showed different coordination patterns in pelvis-to-hip, hip-to-knee, and hip-to-ankle couplings in some parts of the gait cycle. It concludes that the active TTAs with PLs walked with more flexion of the knee and hip, which may indicate a progressive walking strategy and the differences in coordination patterns suggest active TTA individuals used different neuromuscular control strategies to adapt to their amputation. Researchers can extend this work by investigating variations in these parameters across diverse patient populations, including different amputation etiologies and prosthetic designs. Moreover, Clinicians can use the findings to tailor rehabilitation programs for TTAs, emphasizing joint flexibility and coordination.