Background Many gait impairments are characterized by asymmetry and result in reduced mobility. Exoskeletons could be useful for restoring gait symmetry by assisting only one leg. However, we still have limited understanding of the effects of unilateral exoskeleton assistance. Our aim was to compare the effects of unilateral and bilateral assistance using a within-subject study design. Methods Eleven participants walked in different exoskeleton conditions. In the Unilateral conditions, only one leg was assisted. In Bilateral Matched Total Work, half of the assistance from the Unilateral conditions was applied to both legs such that the bilateral sum was equal to that of the Unilateral conditions. In Bilateral Matched Work Per Leg, the same assistance as in the Unilateral conditions was provided to both legs such that the bilateral sum was the double of that of the Unilateral conditions. In the Powered-Off condition, no assistance was provided. We measured metabolic energy consumption, exoskeleton mechanics and kinematics. Results On average, the Unilateral, Bilateral Matched Total Work and Bilateral Matched Work Per Leg conditions reduced the metabolic rate by 7, 11 and 15%, respectively, compared with the Powered-Off condition. A possible explanation for why the Unilateral conditions effectively reduced the metabolic rate could be that they caused only very little asymmetry in gait biomechanics, except at the ankle and in the horizontal center-of-mass velocity. We found the highest ratio of metabolic rate reduction versus positive work assistance with bilateral assistance and low work per leg (Bilateral Matched Total Work). Statistical analysis indicated that assistance symmetry and assistance per leg are more important than the bilateral summed assistance for reducing the metabolic rate of walking. Conclusions These data bridge the gap between conclusions from studies with unilateral and bilateral exoskeletons and inform how unilateral assistance can be used to influence gait parameters, such as center-of-mass velocity. Electronic supplementary material The online version of this article (10.1186/s12984-018-0381-z) contains supplementary material, which is available to authorized users.