| 噬菌体展示EpCAM单链抗体及免疫原性初步研究 |
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| 引用本文： | 刘晓丽,张岸梅,林盛,王欣欣,廖荣霞,孙建国,陈正堂.噬菌体展示EpCAM单链抗体及免疫原性初步研究[J].第三军医大学学报,2011,33(10):977-979. |
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| 作者姓名： | 刘晓丽 张岸梅 林盛 王欣欣 廖荣霞 孙建国 陈正堂 |
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| 作者单位： | 刘晓丽,张岸梅,林盛,王欣欣,孙建国,陈正堂,Liu Xiaoli,Zhang Anmei,Lin Sheng,Wang Xinxin,Sun Jianguo,Chen Zhengtang(第三军医大学新桥医院全军肿瘤研究所,重庆,400037);廖荣霞,Liao Rongxia(第三军医大学基础医学部医学英语教研室,重庆,400038) |
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| 基金项目： | 国家高技术研究发展计划863计划项目，国家自然科学基金 |
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| 摘 要： | 目的利用噬菌体展示技术进行鼠抗人EpCAM单链抗体表达并鉴定其免疫活性,构建特异靶向上皮来源肿瘤细胞及癌干细胞的单链抗体。方法采用全基因合成的方法合成VL+Linker+VH的EpCAM的单链抗体(ScFv)基因,克隆到pMD19-T简易载体中,将ScFv基因酶切并纯化后克隆到噬菌体载体pCANTAB-5E,热休克法转化至感受态的大肠杆菌TG1,随机挑取5个克隆,经辅助噬菌体超感染,形成鼠抗人EpCAM噬菌体单链抗体,用ELISA(以P/N>2判定为阳性)和免疫细胞化学法测定其抗原结合活性。结果成功获得pCANTAB-5E/EpCAM ScFv重组克隆,经ELISA和免疫细胞化学测定,噬菌体展示阳性率为20%,展示的单链抗体能与EpCAM抗原和A549细胞特异性结合。结论噬菌体展示技术制备EpCAM单链抗体方法简便、省时、高效。
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| 关 键 词： | 单链抗体 噬菌体展示 EpCAM 免疫 肿瘤干细胞 |
Preliminary study on immunogenicity of phage-displayed mouse-anti-human EpCAM single-chain antibody |
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| Liu Xiaoli,Zhang Anmei,Lin Sheng,Wang Xinxin,Liao Rongxia,Sun Jianguo,Chen Zhengtang.Preliminary study on immunogenicity of phage-displayed mouse-anti-human EpCAM single-chain antibody[J].Acta Academiae Medicinae Militaris Tertiae,2011,33(10):977-979. |
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| Authors: | Liu Xiaoli Zhang Anmei Lin Sheng Wang Xinxin Liao Rongxia Sun Jianguo Chen Zhengtang |
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| Institution: | Liu Xiaoli1,Zhang Anmei1,Lin Sheng1,Wang Xinxin1,Liao Rongxia2,Sun Jianguo1,Chen Zhengtang1(1Cancer Research Center,Xinqiao Hospital,Third Military Medical University,Chongqing,400037,2Department of Medical English,College of Basic Medical Sciences,400038,China) |
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| Abstract: | Objective To express mouse-anti-human EpCAM single-chain antibody(ScFv) specifically targeting tumor cells or cancer stem cells of epithelial origin via phage display technology,and to identify its immunological activity.Methods Total gene synthesis method was used to synthesize mouse-anti-human EpCAM ScFv gene with a structure of VL+Linker+VH.The gene was cloned to pMD19-T Simple Vector and then was digested,after being purified,was cloned to phage vector pCANTAB 5E.Competent E.coli TG1 cells were transfor... |
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| Keywords: | EpCAM |
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