Experimental studies report that ligaments of the ankle joint are prestrained. The prestrain is an important aspect of modern biomechanical analysis, which can be included in the models by: applying symmetrical, arbitrary prestrains to the ligaments, assuming a strain-free location for the joint or by using experimental prestrain data. The aim of the study was to comparatively analyze these approaches. In total, 4 prestraining methods were considered. In order to do so, a symmetrical model of the ankle with six nonlinear cables and two sphere–sphere contact pairs was assumed. The model was solved in statics under moment loads up to 5 Nm. The obtained results showed that the arbitrary prestrains caused an unbalanced load for the model at rest, and in turn modified its rest location in an unpredictable way. Due to the imbalance, it was impossible to enforce the assumed prestrains and thus cartilage prestrain was required to stabilize the model. The prestraining had a significant effect on the angular displacements and the load state of the model. The findings suggest that the prestrain values are patient specific and arbitrary prestrains will not be valid for most models.