Filtering criteria Included protein bands Bands specificity in immunoblot (%) Number of identified peptides Number of different proteins Percentage sequence coverage (%) Number of discriminant peptide(s) Any protein sequence coverage STa-98 kDa, ST-36 kDa, ST-33 kDa, ST-32 kDa, ST-21 kDa, MTb-41 kDa, MT-35 kDa, MT-32 kDa, MT-30 kDa. any 1933 732 0.25 to 80 1 to 39 Protein sequence coverage ≥ 40%       ST-98 kDa, ST-36 kDa, ST-33 kDa, ST-32 kDa, ST-21 kDa. any 61 46 40 to 80 1 to 39 Protein sequence coverage  ≥ 40% MT-41 kDa, MT-35 kDa, MT-32 kDa, MT-30 kDa. any 33 27 40 to 70 4 to 20 Protein sequence coverage  ≥ 40% ST-98 kDa, ST-36 kDa, ST-33 kDa, ST-32 kDa, ST-21 kDa, MT-41 kDa, MT-35 kDa, MT-32 kDa, MT-30 kDa. any 94 57 40 to 80 1 to 39 Protein sequence coverage ≥ 40% and a closest ortholog ≤ 90% (NCBI non redundant protein sequence database). ST-98 kDa, ST-36 kDa, ST-33 kDa, ST-32 kDa, ST-21 kDa, MT-41 kDa, MT-35 kDa, MT-32 kDa, MT-30 kDa. any 34 13 40 to 80 4 to 21 Protein sequence coverage ≥ 40% and a closest ortholog ≤ 90% (NCBIc non redundant protein sequence database and ToxoDBd). ST-33 kDa, ST-32 kDa, MT-32 kDa, MT-30 kDa. ≥ 90 19 7 41 to 8 4 to 13 aSoluble Tachyzoite antigens, bMembrane Tachyzoite antigens, cNational Center for Biotechnology Information, dToxoplasma gondii genome Database.

Table 3: Filtering criteria applied to nanoLC-MS/MS to identify specific immunogenic proteins of T. gondii.