The influence of various Zr contents (0–45 wt.%) on the microstructure and mechanical properties of Ti6Al4V alloy was investigated through a combinatorial approach. The diffusion multiples of Ti6Al4V–Ti6Al4V20Fe–Ti6Al4V20Cr–Ti6Al4V20Mo–Ti6Al4V45Zr were manufactured and diffusion-annealed to obtain a large composition space. Scanning electron microscopy, electron probe micro-analysis, and a microhardness system were combined to determine the relationships among the composition, microstructure, and hardness of these alloys. The Ti–6Al–4V–30Zr alloy was found to contain the thinnest α lath and showed peak hardness. X-ray diffraction and transmission electron microscope results indicated that after quenching from the β-field, the metastable α″-phase formed; moreover, at the secondary aging stage, the metastable α″-phase acted as precursor nucleation sites for the stable α-phase. The bulk Ti6Al4V30Zr alloy was manufactured. After aging at 550 °C, the alloy showed excellent balance of strength and ductility, and the tensile strength was 1464 MPa with a moderate elongation (8.3%). As the aging temperature increased, the tensile strength and yield strength of the alloys rose, but the total elongation decreased. The lamella thickness and volume fraction of the α-phase were the major factors that had great impacts on the mechanical properties.