Text S1 - Proteomic Selection of Immunodiagnostic Antigens for Trypanosoma congolense

Figure S1. SDS-PAGE with Coomassie blue staining of purified recombinant proteins. The names of the expressed proteins appear above each respective gel lane. The bands indicated by arrow heads are: A, Tc38630; B, degradation product of Tc38630; C, E. coli Ef-Tu (co-purifying contaminant); D, Tc29290. These identities were confirmed by tryptic digestion and mass spectrometry. Figure S2. Assessment of Tc38630, Tc29290 and Tc51750 ELISA assays with pre- and post-infection calf sera. (A) ELISA plates coated with the three recombinant proteins were tested with pre-infection (day −7; n = 40) and post-infection (day +28; n = 40) calf sera. The data were plotted on RoC curves of specificity against selectivity (1-specificity). The output statistics show a sensitivity and specificity of 90% and 94.2% for Tc38630, 80% and 67.3% for Tc29290 and 82.5% and 67.3% for Tc51750, respectively. Table S1. Antigens selectively recognised by T. congolense infection IgG. The antigens are ordered by infection : control LC-MS/MS intensity and coloured coded according to their by LC-MS/MS intensities: black bold >1000, black >60, grey 1000) and an infection : control ratio >100 were assigned identities. Those that failed to yield soluble protein in expression and purification trials are marked with ▪. Those selected but untested in expression trials are marked with white rectangle (vectors available on request) and those purified successfully are marked white rectangle. Table S2. Amino acid sequences of the seven antigen domains successfully expressed in E. coli. Differences in sequence to those in the TriTrypDB are highlighted in red, these are probably due to strain variation. (DOC)

图S1 考马斯亮蓝染色的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果，展示纯化后的重组蛋白。每条凝胶泳道上方标注了对应表达蛋白的名称。箭头指示的条带分别为：A，Tc38630；B，Tc38630的降解产物；C，大肠杆菌（E. coli）延伸因子Tu（共纯化污染物）；D，Tc29290。上述蛋白的身份均通过胰蛋白酶消化与质谱分析得到确认。

图S2 利用感染前后犊牛血清评估Tc38630、Tc29290与Tc51750的酶联免疫吸附实验（ELISA）检测效果。(A) 将三种重组蛋白包被的酶联免疫吸附实验板，分别使用感染前（第-7天；n=40）与感染后（第+28天；n=40）的犊牛血清进行检测。数据以特异性为纵轴、假阳性率（1-特异性）为横轴绘制受试者工作特征（ROC）曲线。输出统计结果显示，Tc38630的灵敏度与特异性分别为90%和94.2%，Tc29290为80%和67.3%，Tc51750为82.5%和67.3%。

表S1 刚果锥虫（T. congolense）感染宿主血清IgG选择性识别的抗原。抗原按感染组与对照组的液相色谱-串联质谱（LC-MS/MS）强度比值排序，并依据LC-MS/MS强度进行颜色编码：黑色粗体代表强度>1000，黑色代表强度>60，灰色代表强度<60；同时，LC-MS/MS强度>1000且感染组/对照组强度比>100的抗原被赋予明确身份。在表达与纯化实验中无法获得可溶性蛋白的抗原以方块（▪）标记；经筛选但未开展表达实验的抗原以白色矩形标记（载体可按需索取）；成功纯化的抗原以白色矩形标记。

表S2 7个在大肠杆菌（E. coli）中成功表达的抗原结构域的氨基酸序列。与TriTrypDB数据库中序列的差异以红色标注，该差异大概率源于菌株变异。（DOC）