用SEREX方法筛选HCC抗原及hcct-19表达谱的检测
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陈晓红, 陈 智, 陈 峰, 姚航平, 夏 琦,
SEREX;肝细胞癌;抗原;hcct-19;表达谱;人组织;细胞株陈晓红,陈智,陈峰,姚航平,夏琦,焦婧婧.用SEREX方法筛选HCC抗原及hcct-19表达谱的检测. 世界华人消化杂志2
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陈晓红, 陈智, 陈峰, 姚航平, 夏琦, 焦婧婧, 浙江大学医学院附属第一医院传染病研究所 浙江省杭州市 310003
陈晓红, 2005年浙江大学内科学博士, 现在哈尔滨医科大学第一临床医学院感染科工作, 主治医师、讲师, 主要从事病毒性肝炎及原发性肝癌发病机制的研究.
通讯作者: 陈智, 310003, 浙江省杭州市庆春路79号, 浙江大学附属第一医院传染病研究所. chenzhi@zju.edu.cn
电话: 0571-87236580
收稿日期: 2005-11-08 接受日期: 2005-12-24
Identification of hepatocellular carcinoma antigens by serological analysis of recombinant cDNA expression library and detection of hcct-19 expression in human tissues and cell lines
Xiao-Hong Chen, Zhi Chen, Feng Chen, Hang-Ping Yao, Qi Xia, Jing-Jing Jiao
Xiao-Hong Chen, Zhi Chen, Feng Chen, Hang-Ping Yao, Qi Xia, Jing-Jing Jiao, Institute of Infectious Diseases, the First Affiliated Hospital of Zhejiang University College of Medicine, Hangzhou 310003, Zhejiang Province, China
Correspondence to: Zhi Chen, Institute of Infectious Diseases, the First Affiliated Hospital of Zhejiang University College of Medicine, 79 Qingchun Street, Hanghzou 310003, Zhejiang Province, China. chenzhi@zju.edu.cn
Received: 2005-11-08 Accepted: 2005-12-24
AbstractAIM:To identify hepatocellular carcinoma (HCC) antigens by serological analysis of recombinant cDNA expression library (SEREX) and observe the expression of hcct-19 in human tissues and cell lines.
METHODS: Firstly, a high-quality cDNA library from human HCC tissue was constructed successfully. E.coli transfected with recombinantλTripleX2 phages were plated onto LB-agar plates. The expression of recombinant proteins was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG). The membranes were blocked with BSA and incubated with a 1∶1000 dilution of patient’s serum, which had been preabsorbed with transfectedE.coli phage lysate. Then they were incubated with a 1∶5000 dilution of the alkaline phosphatase-conjugated antibody specific for human IgG. Reactive clones were visualized by staining with BCIP/NBT, and these clones were subcloned three times to obtain monoclonality ......
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