| 用EBV转化的肝癌患者B细胞构建噬菌体单链抗体库 |
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| 引用本文： | 肖艳,李官成,李跃辉,童永清,张志杰.用EBV转化的肝癌患者B细胞构建噬菌体单链抗体库[J].中国现代医学杂志,2008,18(8):1040-1044. |
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| 作者姓名： | 肖艳 李官成 李跃辉 童永清 张志杰 |
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| 作者单位： | 中南大学肿瘤研究所,免疫室,湖南,长沙,410078 |
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| 摘 要： | 目的 构建抗肝癌人源噬菌体单链抗体库.方法 体外致敏并用EBV转化肝癌患者的PBMC.用PCR分别扩增VH和VL基因并组成ScFv基因.将ScFv基因与载体连接后,转化大肠杆菌MC1061,构建噬菌体呈现型ScFv库.结果 经EBV转化的4例肝癌患者PBMC,ELISA检测均有抗肝癌抗体产生,经多次PCK,扩增出6种vH(γ、μ)和9种VL(κ、λ)基因,经连接组成54种ScFv基因.将ScFv基因与载体连接后.导入大肠杆菌MC1 061.经四环素抗性筛选,得到库容为1.0×108的初级噬菌体抗独特型抗体库,噬菌体DNA中全长ScFv基因的插入率为80%.结论 用体外致敏法结合噬菌体抗体库技术,制备全人源抗肿瘤单链抗体(ScFv),这一策略是完全可行的.
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| 关 键 词： | EB病毒转化 噬菌体抗体库 肝癌 单链抗体 转化大肠杆菌 肝癌患者 细胞构建 噬菌体抗体库技术 单链抗体库 cancer patients liver in vitro antibody library phage 完全 策略 抗肿瘤 结合 抗独特型 库容 抗性筛选 四环素 抗体产生 检测 |
| 文章编号： | 1005-8982(2008)08-1040-05 |
| 修稿时间： | 2007年10月9日 |
Construction of phage antibody library using sensitized in vitro B-lymphocytes of liver cancer patients |
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| XIAO Yan,LI Guan-cheng,LI Yue-hui,TONG Yong-qing,ZHANG Zhi-jie.Construction of phage antibody library using sensitized in vitro B-lymphocytes of liver cancer patients[J].China Journal of Modern Medicine,2008,18(8):1040-1044. |
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| Authors: | XIAO Yan LI Guan-cheng LI Yue-hui TONG Yong-qing ZHANG Zhi-jie |
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| Institution: | XIAO Yan,LI Guan-cheng,LI Yue-hui,TONG Yong-qing,ZHANG Zhi-jie (Cancer Research Institute of Central South University,Changsha,Hunan 410078,P.R.China) |
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| Abstract: | Objective To construct fully humanized phage antibody library. Methods Peripheral blood mononuclear cells (PBMCs) of patients with liver cancer were sensitized in vitro and transformed by Epstein-Barr virus (EBV). VH and VL genes were reamplified by PCR and combined to single-chain fragment of variable region (ScFv) genes. ScFv genes were cloned into vector FUSE5 and transformed into MC1 061 by electroporation to construct the ScFv-displaying phage library. Results Detection of ELISA showed that 2 liver can... |
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| Keywords: | EBV immortalization phage antibody libraries liver cancer single-chain Fv |
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