双向电泳联合免疫印迹技术分析日本血吸虫成虫可溶性抗原  

Analysis on the Soluble Antigen of Schistosoma Japonicum Adult Worm with Two Dimensional Electrophoresis Followed by Immunoblotting

作　　者：郑辉[1,2] 吴赟[2] 余轶婧[1] 钟政荣[1] 罗庆礼[1] 沈继龙[1] 

机构地区：[1]安徽医科大学人畜共患病安徽省重点实验室,重要遗传病基因资源利用教育部省部共建重点实验室,安徽医科大学病原生物学教研室,合肥230032 [2]安徽医科大学附属省立医院检验科

出　　处：《临床输血与检验》2009年第2期107-112,共6页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：国家高技术研究发展计划(863计划)项目(No.2004AA2Z3570)资助

摘　　要：目的建立及优化日本血吸虫成虫的蛋白质组学分析方法,寻找可溶性蛋白组分中与免疫应答相关的特异性成虫抗原。方法纯化日本血吸虫成虫可溶性蛋白,应用双向电泳(2D-E)结合免疫印迹技术(Western blotting),获得相应的电泳图谱和免疫印迹图谱,应用PDQuest8.0双向电泳图像分析软件对图像进行分析比对,鉴别特异性抗原。结果血吸虫成虫可溶性蛋白经双向电泳后,考马斯亮蓝G250染色,电泳图谱显示约152个主要蛋白斑点。分子量(Mr)分布约为14～114kD;等电点(pI)主要集中在4.9～9.5。进一步的Western blotting鉴定结果显示:实验组图像可见的抗原抗体反应点数目约57个,对照组为0。结论双向电泳联合免疫印迹技术是成功分析蛋白质组学的技术关键,该技术为寻找日本血吸虫特异性抗原开辟了新途径。Objective To establish and optimize the proteomic analysis of schistosoma japonicum adult worm and to provide basic foundation for the identification of the specific antigens in soluble proteins of schistosoma japonicum adult worm for further research. Methods To employ the techniques of two-dimensional eleetrophoresis （2-DE） combined with immunoblotting assay（western blotting） and to analyze the soluble components of the adult worm antigens （AWA） of schistosoma japonicum. Acquired 2-DE map and immunoblotting map were analysed with the image analysis software （PDQuest 8.0）. Results About 152 soluble proteins spot.4 ＇were revealed in eoomassie-stained gels. Most of the proteins had a molecular weight （Mr）of 14 000 to 114 000, and an isoelectric point （pI）value of 4.9 to 9.5. Immunoblotting of 2- DE were showed and there were 57 specific antigen spots, compared with the controls. Conclusion The technique of 2-DE combined with western blotting is the key for the successful analysis of the proteomics, which presents a new possibility for searching the specific antigen of schistosoma japonicum.

关 键 词：日本血吸虫 蛋白质组学 成虫特异性抗原 双向电泳 免疫印迹 

分 类 号：R532.21[医药卫生—内科学] R446.61[医药卫生—临床医学]