人肝癌血清对人体胚胎干细胞文库的筛选及方法改进  

Screen cDNA Expression Library of Human Embryonic Stem Cells by Sera of HCC and the Methods Modified

作　　者：孙泽峰[1] 郭雪婷[1] 李雅娟[1] 程学远[1] 周素芳[1] 

机构地区：[1]广西医科大学生物化学与分子生物学教研室,南宁530021

出　　处：《生物技术通报》2008年第S1期160-164,共5页Biotechnology Bulletin

基　　金：广西区研究生创新课题资助项目(2006105981001M01);国家自然科学基金项目(30760233);广西自然科学基金项目(0728121)

摘　　要：用人肝癌血清从人体胚胎干细胞cDNA表达文库中筛选肝癌相关抗原,并对SEREX条件方法的改良作进一步的探索。将已构建好的人胚胎干细胞cDNA表达文库进行滴度计算。利用38例原发性肝癌病人血清,结合SEREX技术,应用Western blot方法,对表达文库中的相关抗原做血清学免疫筛选。同时,对SEREX技术贴膜条件及假阳性的排除方法进行改良。cDNA表达文库滴度为3.8×10~8pfu/ml,筛选出阳性克隆有31个。从人胚胎干细胞cDNA表达文库中能筛查到部分有效的肝癌肿瘤相关抗原,为后续工作提供宝贵资料。To find out antigenic genes of human hepatocellular carcinoma(HCC)from cDNA expression librarie of human embryonic stem cells.to explore methods modified of SEREX.Methods the titer of a cDNA expression library of human embryonic stem cells was determinated.The recombinant cDNA expression library was screened with sera of HCC by the methods of SEREX and Westernblot.Besides,we improve experimental condition to lower fake postive.Results the titer of cDNA expression library was 3.8×10~8 pfu/ml recombinants.31 antigenic genes were identified.Conclusion some hepatocellular carcinoma-associated antigen can be screened by the method of SEREX.these antigens will provide important data for the future research.

关 键 词：人体胚胎干细胞cDNA文库 SEREX技术 肝癌相关抗原 

分 类 号：R735.7[医药卫生—肿瘤]