Background: This study aimed to determine preoperative predictions of sagittal imbalance in kyphosis secondary to ankylosing spondylitis (AS) after one-level three-column osteotomy. Methods: A total of 55 patients with AS who underwent one-level three-column osteotomy were enrolled. The patients were divided into two groups according to sagittal vertical axis (SVA) value at the final follow-up (group A: SVA>5 cm; group B: SVA≤5 cm). The radiographic measurements were measured, including global kyphosis, lumbar lordosis (LL), pelvic tilt (PT), pelvic incidence (PI), sacral slope, T1 pelvic angle (TPA), SVA, osteotomized vertebral angle and PI and LL mismatch (PI-LL). Results: Fifty-five AS patients were followed up for 30.6 ± 10.2 months averagely (range, 24–84 months). Preoperatively and postoperatively, group A had larger LL, PT, PI - LL, TPA, and SVA values than group B (P<0.05). And the preoperative LL, PT, PI - LL, TPA, and SVA values had significantly positive relationships with the follow-up SVA value (P<0.05). The preoperative TPA>40.9°, PI-LL>32.5°, and SVA>13.7 cm were the top three predictions with the best accuracy to predict sagittal imbalance. The construction of postoperative SVA<7.4 cm was the key factor for decreasing the chance of sagittal imbalance on follow-up. Conclusions: The preoperative TPA>40.9°, PI - LL>32.5°, and SVA>13.7 cm could predict sagittal imbalance for AS kyphosis after one-level three-column osteotomy, and additional osteotomies were recommended for this condition. The postoperative SVA<7.4 cm was an optimal alignment for preventing sagittal imbalance. Level of evidence: IV