One of the most significant developments in prosthetic knee technology has been the introduction of the Microprocessor-Controlled Prosthetic Knee (MPK). However, there is a lack of consensus over how different types of MPKs affect performance in different ambulation modes. In this study, we investigated the biomechanical differences in ramp and stair maneuvers when an individual with transfemoral amputation wears three commercial MPKs: the Össur Power Knee, the Össur Rheo Knee and the Ottobock C-Leg 4. The primary outcome variable for this study was the lower limb biological joint work, inclusive of the intact leg and prosthetic side hip. We hypothesized that (1) the Power Knee would result in lower biological work during ascent activities than the C-Leg and Rheo, both passive MPKs, and (2) the C-Leg and Rheo would result in lower biological work during descent activities than the Power Knee. During ramp ascent, the C-Leg was associated with lower biological joint work (p < 0.05) than the Power Knee. However, this relationship did not hold during stair ascent, where the Power Knee showed advantages for stair ascent with net reductions in biological joint work of 14.1% and 23.3% compared to the Rheo and C-leg, respectively. There were no significant differences in biological joint work between the knees during ramp and stair descent, indicating that choice of MPK may not be as important for descent activities. Our results demonstrate that differences are present between different types of MPKs during ascent activities which could prove useful in the prescription of these devices.