Isocyanate-based modification is lately gaining acceptance as a successful way to give added value to bitumen, a crude oil refining by-product. In order to study the influence of prepolymer type on the rheological properties of the resulting binders, six prepolymers synthesized from polypropylene-glycols (PPG) with varying molecular weight (between 440 and 2425) and different molar excess of a polymeric MDI (4,4'-diphenylmethane diisocyanate) were used. Two modification procedures, either involving or not water addition were followed. The modification achieved depends on both the selected polyol molecular weight and the excess in MDI (i.e., free isocyanate content), although not in a similar extent. Viscous flow and dynamic oscillatory shear tests, at 60 ºC, demonstrated a much higher level of bitumen modification by using the prepolymer prepared with the polyol having a molecular weight of 940 and with a free isocyanate content of 17.4 wt.%, mainly after addition of water. On the other hand, bitumen nature greatly affects the final rheological properties of these bituminous products. In that sense, modification results much more effective when conducted on bitumen with a well-developed colloidal microstructure.