| 抗人肝癌单链抗体的基因克隆和分泌型表达 |
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| 引用本文： | 牛利国,陈志南.抗人肝癌单链抗体的基因克隆和分泌型表达[J].细胞与分子免疫学杂志,1998,14(2):102-104,109. |
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| 作者姓名： | 牛利国 陈志南 |
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| 作者单位： | 第四军医大学生物化学及分子生物学教研室 |
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| 摘 要： | 应用ＲＴ－ＰＣＲ技术，从分泌抗人肝癌单克隆抗体的杂交瘤细胞ＨＡｂ１８中，扩增出抗体ＶＨ和ＶＬ连接肽基因，将ＶＨ和ＶＬ连接成ＳｃＦｖ基因，并进行序列测定，结果表明，ＶＨ，ＶＬ，ｌｉｎｋｅｒ拼接正确，基因全长为７２６ｂｐ。用噬菌粒表达载体ｐＣＡＮＴＡＢ ５Ｅ在大肠杆菌中表达了可溶性的ＳcＦｖ融合蛋白，经流式细胞仪分析表达产物可特异地与肝癌细胞结合，而不与正常肝细胞及胃癌细胞结合。
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| 关 键 词： | 单链抗体 克隆 表达 噬菌粒载体 肝肿瘤 |
Cloning and expression of ScFv gene for an anti human liver cancer antibody |
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| NIU Li Guo,CHEN Zhi Nan,SU Cheng Zhi,YAO Li Bo,LIU Xin Ping,WANG Guo Hua,YANG Min.Cloning and expression of ScFv gene for an anti human liver cancer antibody[J].Journal of Cellular and Molecular Immunology,1998,14(2):102-104,109. |
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| Authors: | NIU Li Guo CHEN Zhi Nan SU Cheng Zhi YAO Li Bo LIU Xin Ping WANG Guo Hua YANG Min |
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| Abstract: | The V H and V L genes were amplified from a hybridoma cell line HAb18 producing mouse mAb against human liver cancer by RT PCR. The genes were sequenced and compared with those published genes in EMBL Genbank. The results showed that the V H and V L genes were homologous with the published gene sequences of mouse antibody variable region. The V H and V L genes were connected through a flexible linker(Gly 4Ser) 3, and the V H linker V L(ScFv) fusion gene was cloned into a phagemid vector pCANTAB 5E and expressed in E.coli HB2151. Immunofluorescence assay showed that the expressed protein could bind to human liver cancer cell line SMMC 7721, but not to normal liver cell line QZG and gastric cancer cell line SCG 7901. |
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| Keywords: | ScFv cloning expression liver cancer phagemid vector |
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