Analysis of Protoscoleces-specific Antigens from Echinococcus Granulosus with Proteomics Combined with Western Blot
Analysis of Protoscoleces-specific Antigens from Echinococcus Granulosus with Proteomics Combined with Western Blot
复制标题
蛋白质组学结合Western Blot分析细粒棘球绦虫原头节特异性抗原
DOI：
10.3967/0895-3988.2012.06.015
复制
发表时间：
2012-12-01
影响因子：
3.5
通讯作者：
Zhao Wei
中科院分区：
文献类型：
--
作者：
Li Zong Ji;Zhao Wei
目的建立并优化细粒棘球绦虫原头节特异抗原的蛋白质组学分析方法。目的为进一步研究细粒棘球蚴原头节可溶性蛋白特异性抗原提供基础。方法对1例感染大肠杆菌患者肝脏可育性颗粒绦虫囊进行无菌收集。破碎可育的颗粒棘球蚴囊，离心收集原头节。根据制造商的说明，使用2D定量试剂盒获得原头节的可溶性蛋白。采用双向电泳（2-DE）结合免疫印迹法（Western blot）对颗粒棘球蚴原头节抗原的可溶性成分进行了分析。用PDQuest 8.0图像分析软件分析得到的2-DE图谱和免疫印迹图谱。结果comasse染色凝胶共鉴定出233个可溶性蛋白点。大部分蛋白分子量在16000 ~ 117000 Da之间，等电点值在3.0 ~ 10.0之间。2-DE免疫印迹法观察到57个特异性抗原斑点，其中鉴定出23个特异性抗原斑点。结论2-DE联合Western blot是成功进行蛋白质组学分析的关键，为寻找颗粒棘球蚴原头节特异性抗原提供了新的可能性。
Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research.Methods Brood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer's instructions. We employed two-dimensional electrophoresis (2-DE) combined with immunoblot assay (Western blot) to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software.Results About 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16 000 Da to 117 000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified.Conclusion 2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.