In the present study, using two-dimensional gel electrophoresis (2DE), mice sera immunoblotting, and mass spectrometry (MS) analysis, we attempted to discover three previously uncharacterized antigens of T. gondii, EF1γ, PGKI, and GAP50—which have the potential to distinctly target T. gondii IgM antibodies during the early infection of rodent model. Here, CD40LG is linked to infection.