| 胰腺癌人源噬菌体单链抗体库的构建及初步筛选 |
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| 引用本文： | 何光志,田维毅,高英,王平,王文佳,黄高,蔡琨,安传伟.胰腺癌人源噬菌体单链抗体库的构建及初步筛选[J].广东农业科学,2011,38(7):158-160. |
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| 作者姓名： | 何光志 田维毅 高英 王平 王文佳 黄高 蔡琨 安传伟 |
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| 作者单位： | 贵阳中医学院;遵义医学院附院; |
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| 基金项目： | 贵州省科学技术基金(黔科合J字[2010]2262号) |
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| 摘 要： | 提取胰腺癌患者的外周淋巴细胞总RNA,以RT-PCR技术分别扩增出人源抗体H链和L链的可变区基因,采用SOE-PCR法将VH和VL片段随机拼接成scFv片段,然后将scFv片段克隆至pCANTAB5E载体,并电转化至感受态TG1菌,经辅助噬菌体M13K07拯救,获得胰腺癌人源噬菌体单链抗体库。结果表明,从外周血淋巴细胞中获取可变区基因,利用噬菌体抗体库技术制备人源抗胰腺癌单链抗体的策略是可行的。
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| 关 键 词： | 胰腺癌 人源噬菌体单链抗体 构建与筛选 |
Construction and screening of single-chain antibody library of pancreatic cancer |
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| HE Guang-zhi,TIAN Wei-yi,GAO Ying,WANG Ping,WANG Wen-jia,HUANG Gao,CAI Kun,AN Chuan-wei.Construction and screening of single-chain antibody library of pancreatic cancer[J].Guangdong Agricultural Sciences,2011,38(7):158-160. |
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| Authors: | HE Guang-zhi TIAN Wei-yi GAO Ying WANG Ping WANG Wen-jia HUANG Gao CAI Kun AN Chuan-wei |
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| Affiliation: | HE Guang-zhi1,TIAN Wei-yi1,GAO Ying2,WANG Ping 1,WANG Wen-jia1,HUANG Gao1,CAI Kun1,AN Chuan-wei 1(1.Guiyang College of Traditional Chinese Medicine,Guiyang 550002,China,2.Affiliated Hospital of Zunyi Medical College,Zunyi 563003,China) |
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| Abstract: | Total RNA was abstracted from peripheral blood lymphocytes of patients suffering from pancreatic cancer,and was used to amplify VH and VL gene by RT-PCR.VH and VL were joined by SOE-PCR to form the single chain variable fragment(ScFv) gene.These ScFv fragments were cloned into the phagemid pCANTAB5E and the recombinant phagemids were ransformed to susceptible E.coli TGl by roporation.After rescuing by helper phage M13K07,a natural single-chain antibody library for pancreatic cancer was successfully construc... |
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| Keywords: | pancreatic cancer human phage antibody construction and screening |
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