In this study, we used Two-dimensional gel electrophoresis (2-DE)-based Western blotting coupled with MALDI-TOF / TOF-MS / MS to separate and identify the T. pseudospiralis adult worms (Ad) ES products immunoreaction-positive proteins. In total, 400 protein spots were separated by 2-DE. 28 protein spots were successfully identified using the sera from infected pigs and were characterized to correlate with twelve different proteins of T. pseudospiralis.