Electron beam melting (EBM) is a layer by layer additive manufacturing technology, which has the capability of producing near-net shaped parts with complex geometries. It is also suitable for handling high melting point and reactive metallic materials, such as Ti alloy, which is widely used in the aerospace and biomedical applications. The present study focused on the relationship between the microstructure and mechanical properties of big-sized Ti-6Al-4V parts. A plate (6mm×180mm×372mm) was additively manufactured by EBM. The microstructure evolution and variation of mechanical properties were investigated by using the x-ray diffraction, optical microscope, scanning electron microscope and tensile test. The results revealed that with an increasing in the build height, there was a variation in the microstructure and the mechanical properties of the build plate. Although only α phase and a relatively small fraction of β phase were detected in both the bottom and top specimens of the build plate, yield strength and ultimate tensile strength decreased with an increase of build height. This was attributed to the increase of α lath width which was caused by the different thermal histories along the build height of the plate.