METHODS Human Cadaveric Biomechanical Study. OBJECTIVE Lumbar Lateral Interbody Fusion (LLIF) utilizing a wide cage has been reported as having favorable biomechanical characteristics. We examine the biomechanical stability of unilateral pedicle screw and rod fixation after multilevel LLIF utilizing 26 mm wide cages compared to bilateral fixation. METHODS Eight human cadaveric specimens of L1-L5 were included. Specimens were attached to a universal testing machine (MTS 30/G). Three-dimensional specimen range of motion (ROM) was recorded using an optical motion-tracking device. Specimens were tested in 3 conditions: 1) intact, 2) L1-L5 LLIF (4 levels) with unilateral rod, 3) L1-L5 LLIF with bilateral rods. RESULTS From the intact condition, LLIF with unilateral rod decreased flexion-extension by 77%, lateral bending by 53%, and axial rotation by 26%. In LLIF with bilateral rods, flexion-extension decreased by 83%, lateral bending by 64%, and axial rotation by 34%. Comparing unilateral and bilateral fixation, LLIF with bilateral rods reduced ROM by a further 23% in flexion-extension, 25% in lateral bending, and 11% in axial rotation. The difference was statistically significant in flexion-extension and lateral bending (P < .005). CONCLUSIONS Considerable decreases in ROM were observed after multilevel (4-level) LLIF utilizing 26 mm cages supplemented with both unilateral and bilateral pedicle screws and rods. The addition of bilateral fixation provides a 10-25% additional decrease in ROM. These results can inform surgeons of the incremental biomechanical benefit when considering unilateral or bilateral posterior fixation after multilevel LLIF.