Abnormal sacral slope (SS) has shown to increase progression of spondylolisthesis, yet there exists a paucity in biomechanical studies investigating its role in the correction of adult spinal deformity, its influence on lumbosacral shear, and its impact on the instrumentation selection process. This in vitro study investigates the effect of SS on 3 anterior lumbar interbody fusion constructs in a biomechanics laboratory. Nine healthy, fresh-frozen, intact human lumbosacral vertebral segments were tested by applying a 550-N axial load to specimens with an initial SS of 20° on an MTS Bionix test system. Testing was repeated as SS was increased to 50°, in 10° increments, through an angulated testing fixture. Specimens were instrumented using a standalone integrated spacer with self-contained screws (SA), an interbody spacer with posterior pedicle screws (PPS), and an interbody spacer with anterior tension band plate (ATB) in a randomized order. Stiffness was calculated from the linear portion of the load-deformation curve. Ultimate strength was also recorded on the final construct of all specimens (n = 3 per construct) with SS of 40°. Axial stiffness (N/mm) of the L5-S1 motion segment was measured at various angles of SS: for SA 292.9 ± 142.8 (20°), 277.2 ± 113.7 (30°), 237.0 ± 108.7 (40°), 170.3 ± 74.1 (50°); for PPS 371.2 ± 237.5 (20°), 319.8 ± 167.2 (30°), 280.4 ± 151.7 (40°), 233.0 ± 117.6 (50°); and for ATB 323.9 ± 210.4 (20°), 307.8 ± 125.4 (30°), 249.4 ± 126.7 (40°), 217.7 ± 99.4 (50°). Axial compression across the disc space decreased with increasing SS, indicating that SS beyond 40° threshold shifted L5-S1 motion into pure shear, instead of compression-shear, defining a threshold. Trends in ultimate load and displacement differed from linear stiffness with SA > PPS > ATB. At larger SSs, bilateral pedicle screw constructs with spacers were the most stable; however, none of the constructs were significantly stiffer than intact segments. For load to failure, the integrated spacer performed the best; this may be due to angulations of integrated plate screws. Increasing SS significantly reduced stiffness, which indicates that surgeons need to consider using more aggressive fixation techniques.