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目的 分析鉴定日本血吸虫未成熟卵可溶性抗原 (SIEA) 2 8k Da抗原组分。方法 用高效液相离子交换法对电泳纯 SIEA2 8k Da抗原组分进一步分离纯化 ,对获得的色谱纯 SIEA2 8k Da抗原组分进行二维凝胶电泳分析。结果 SIEA经双向凝胶电泳分离、银染后获得的 SIEA- 2 D图谱蛋白斑点以 p I3～ 7和 Mw14～ 6 6 k Da范围最多。进一步经图象采集、PDQuest2 D软件分析 SIEA双向电泳图谱 ,计算在相同的设定值条件下获取的蛋白质斑点数 ,测得 SIEA- 2 D电泳图谱平均为 (94 8±89)个蛋白质斑点数。电泳纯 SIEA2 8k Da抗原组分用高效液相离子交换法纯化后 ,2 80、2 2 0 nm波长色谱图均为一个两侧对称、光滑的高峰曲线 ,色谱洗脱液对其活性光密度图也为一个两侧对称、光滑的高峰曲线。二维凝胶电泳银染色可见一个显色饱和度高的蛋白质圆斑 ,免疫印迹分析为一个显色饱和度高的单一圆斑。结论 色谱纯 SIEA2 8k Da抗原组分为单一分子 ,其等电点为 5 .7
Objective To identify and identify the 28 kDa antigens of immature egg soluble antigen (SIEA) of Schistosoma japonicum. Methods The pure SIEA2 8k Da antigen component was further separated and purified by high performance liquid chromatography (HPLC). The chromatographic pure SIEA2 8k Da antigen component was analyzed by two dimensional gel electrophoresis. Results SIEA was separated by two-dimensional gel electrophoresis. The spots of SIEA-2 D spectra obtained by silver staining were most abundant in pI3 ~ 7 and Mw14 ~ 6 6 kDa. Further image acquisition, PDQuest2 D software SIEA two-dimensional electrophoresis analysis, calculated under the same set of conditions to obtain protein spots, measured SIEA-2 D electrophoresis average (94 8 ± 89) protein spots . Electrophoresis pure SIEA2 8k Da antigen components purified by high performance liquid chromatography after ion exchange, 2 80,220 nm wavelength chromatograms are a bilateral symmetrical smooth peak curve, the chromatographic eluate on the activity of optical density It is also a symmetrical, smooth peak curve on both sides. Two-dimensional gel electrophoresis silver staining shows a high color saturation protein spots, immunoblot analysis of a high color saturation of a single circular spot. Conclusion The chromatographic pure SIEA2 8k Da antigen has a single molecule with an isoelectric point of 5.7