The Toxoplasma gondii cytoskeleton is a highly organized structure essential for parasite motility, replication, and host cell invasion. To identify its components, a highly enriched fraction of tachyzoite cytoskeletons was obtained and quantitatively analyzed by mass spectrometry (LC/ESI/HDMSE). We identified 623 proteins classified into 18 functional groups, including 21 IMC proteins, 26 cytoskeleton proteins, and 42 uncharacterized proteins. A comprehensive bioinformatic analysis was conducted to assess protein abundance (fmol), antigenicity, accessibility, interactome, and homology, with the aim of identifying immunogenic targets. Among the top vaccine candidates were GRA12, IMC1, ROP8, and IMC4, with ROP8 emerging as the most promising based on epitope prediction.