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Titanium-tantalum (Ti-Ta) alloy has excellent biomechanical properties with high strength and low Young’s modulus, showing great application potential in the biomedical industry. In this study, Ti-Ta alloy samples were prepared by laser powder bed fusion (LPBF) technology with mixed pure 75 wt.% Ti and 25 wt.% Ta powders as the feedstock. The maximum relative density of Ti-Ta samples prepared by LPBF reached 99.9%. It is well-accepted that four nonequilibrium phases, namely, α′, α″ and metastable β phase exist in Ti-Ta alloys. The structure of α′, α″ and β are hexagonal close-packed (HCP), base-centered orthorhombic (BCO) and body-centered cubic (BCC), respectively. X-ray Diffraction (XRD) analysis showed that the α′ phase transformed to the α″ phase with the increase of energy density. The lamellar α′/α″ phases and the α″ twins were generated in the prior β phase. The microstructure and mechanical properties of the Ti-Ta alloy were optimized with different LPBF processing parameters. The samples prepared by LPBF energy density of 381 J/mm³ had a favorable ultimate strength (UTS) of 1076 ± 2 MPa and yield strength of 795 ± 16 MPa. The samples prepared by LPBF energy density of 76 had excellent ductility, with an elongation of 31% at fracture.