摘要
目的:建立一种新的分离纯化人肝脏特异性F抗原(肝脏F蛋白)的方法,为临床检测肝脏特异性F抗原提供物质基础。方法:在0.25mol/L蔗糖中,人肝组织匀浆以105000g离心后,上清液用Sepharose6-B和DEAE-ED52两步柱层析分离,最后经等电聚焦,得到纯化的F抗原。用纯化的F抗原免疫兔子获相应的多克隆抗体。结果:纯化的肝脏F抗原经SDS-PAGE电泳鉴定呈单一区带,分子量约44000Da,PI在6.5-6.7之间,密度为1.26,是一种蛋白质抗原,不含有可探测到的碳水化合物和脂类。结论:经两步柱层析及等电聚焦可获得纯化的F抗原,并保留了免疫特性,可供进一步研究其临床价值。
Objective：To establish a new purification method of human liver specific F antigen in order to provide a specific human liver F antigen for clinical diagnosis. Methods：0.25mol/L sucrose contained human liver homogenates was centrifuged at 105000g for 60 min. The supernatants were chromatographied on Sepharose 6-B and then sbujected to DEAE-cellulose column. Finally, purified Fantigen was obtained after preparative isoelectric focusing. The multiple clone antibodies of the F antigen were obtained by immunized of rabbit with purified human liver specific F antigen. Results：This purified human liver-specific F antigen was confirmed by SDS-PAGE gel, which had a band with molecular mass of approximately 44, 000 Daltons,a PI of 6.5-6.7 and a density of 1.26. This protein was an antigen and contained no detectable carbohydrate or lipid. Conclusion： Sepharose 6-B and DEAE-cellulose column combining chromatographied step and isoelectric focusing method provide a purified human liver F antigen, meanwhile, remained its immunological character. The purified human liver F antigen will provide a useful tool for further clinical research.
出处
《温州医学院学报》
CAS
2006年第6期530-533,共4页
Journal of Wenzhou Medical College
关键词
肝脏/F抗原
柱层析
提纯
鉴定
liver/F antigen
column chromatograppy
purification
identification
