| 抗癌胚抗原单区抗体的质粒构建及表达 |
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| 引用本文： | 李彪,吴祥甫.抗癌胚抗原单区抗体的质粒构建及表达[J].上海交通大学学报(医学版),1999,19(2):99-102,109. |
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| 作者姓名： | 李彪 吴祥甫 |
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| 作者单位： | [1]上海第二医科大学附属瑞金医院核医学科 [2]中科院上海生物所 |
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| 基金项目： | 国家自然科学基金!39770231，上海市启明星计划资助项目!98QB14011 |
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| 摘 要： | 目的 对抗癌胚抗原单区抗体进行表达研究。方法 采用ＰＣＲ技术将已获得的抗癌胚抗原（ＣＥＡ）原单克隆抗体Ｅ７Ｂ１０重，轻链可变区基因（ＶＨ、ＶＫ）进行改造后分别克陲是大肠杆菌表达质粒ＰＥＴ－２４α（＋）中，并进行表达，结果 显示重、轻链可变区在大肠杆菌中获得高铲表达，基力体总蛋白的２０％与２５％，ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ ｂｌｏｔ图谱显示表达产物分子量为１３ＫＤ、１２ＫＤ，与其基因编旧白 理
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| 关 键 词： | 癌胚抗原 PCR 基因表达 肿瘤 诊断 治疗 抗体 |
Research on Anti-carcinoembryonic Antigen Single Domain Antibody of Plasmid Construction and Expression |
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| Li Biao, Zhu Chengmo.Research on Anti-carcinoembryonic Antigen Single Domain Antibody of Plasmid Construction and Expression[J].Journal of Shanghai Jiaotong University:Medical Science,1999,19(2):99-102,109. |
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| Authors: | Li Biao Zhu Chengmo |
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| Abstract: | Objective To probe into expression of single domain antibody for anticarcinoembryonic antigen (CEA). Methods The heavy and light chain variable region (VH and VK) genes encoding the murine monoclonal antibody E7B10 with specificity for CEA were amplified by PCR and cloned into expression vector PET-24a( ) by gene recombinant. Results The constructs were expressed in E.Coli with high level as inclusion bodies(IB), reaching 20% and 25% of the total bacteria protein, and characterized by SDS-PAGE and Western blot to have molecula weights of 13KD and 12KD. Conclusion The expression product of the heavy chain variable region has CEA - binding ability in RIA by means of IB solubilization then refolding |
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| Keywords: | carcinoembryonic antigen heavy chain variable region gene light chain variable region gene PCR gene expression |
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