| 乙肝表面抗原专一的单链嵌合抗体在大肠杆菌中的表达 |
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| 引用本文： | 潘华,杨冠珍.乙肝表面抗原专一的单链嵌合抗体在大肠杆菌中的表达[J].遗传学报,1999,26(1):87-91. |
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| 作者姓名： | 潘华 杨冠珍 |
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| 作者单位： | 中科院上海生物化学研究所!上海200031 |
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| 摘 要： | 利用重组ＰＣＲ技术将乙肝表面抗原专一单抗的Ｖｋ与人源的Ｃ基因拼接成轻链嵌合抗体Ｖ－Ｃ，再通过编码柔性肽段（Ｇｌｙ４Ｓｅｒ）的Ｌｉｎｋｅｒ与Ｖ连接成单链嵌合抗体ＳｃＦＶ－Ｃ，并分别在大肠杆菌的热敏与分泌型表达系统中进行表达。表达产物的Ｗｅｓｔｅｒｎ－ｂｌｏｔ与间接ＥＬＩＳＡ检测结果表明，ＳＣＦｖ－Ｃ产两种系统中的表达产物均具有抗原结合活性，并已在分泌肽的指导下ＳｃＦｖ－Ｃｋ能被Ｅ．ｃｏｌｉ分泌出胞外。
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| 关 键 词： | 乙肝表面抗原 单链嵌合抗体 分泌表达 |
Expression of Chimeric Single-Chain Antibody with Specificity for HBsAg in E. coli |
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| PAN Hua, YANG Guan-Zhen, WU Xiang-Fu.Expression of Chimeric Single-Chain Antibody with Specificity for HBsAg in E. coli[J].Journal of Genetics and Genomics,1999,26(1):87-91. |
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| Authors: | PAN Hua YANG Guan-Zhen WU Xiang-Fu |
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| Institution: | Shanghai Institute of Biochemistry Chinese Academy of Sciences Shanghai 200031 |
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| Abstract: | The Vk gene of MAb with specificity for HBsAg and human Ck gene have been combined to form chimeric light chain (Vk-Ck) by recombinant PCR, which has combined with VH, to construct chimeric single-chain antibody(ScFv C,) by a linker encoding a flexible peptide(Gly3 Ser)3]. The ScFv-Ck has been expressed in the heat-induced expression system and secreted expression system of E. coli respectively. Analysis by Western blot and indirect ELISA shows that the ScFv-Ck product in two systems both have HBsAg-binding ability, and ScFv-Ck has been secreted from E. colis in the direction of secretion peptide. |
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| Keywords: | HBsAg ScFV-Ck Heat-induced expression Secreted expression |
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