| IgM型单克隆抗体之scFv库的建立与筛选 |
| |
| 引用本文： | 单超,李兰,王宗华.IgM型单克隆抗体之scFv库的建立与筛选[J].亚热带农业研究,2009,5(2). |
| |
| 作者姓名： | 单超 李兰 王宗华 |
| |
| 作者单位： | 福建农林大学生命科学学院,福建,福州,350002 |
| |
| 基金项目： | 福建省教育厅重点项目 |
| |
| 摘 要： | 从一个抗脂磷酸聚糖IgM型单克隆抗体细胞的mRNA中经RT-PCR克隆重链可变区(VH)和轻链可变区(VL),然后将VH、Linker和VL连接成scFv,并克隆到载体pCANTAB-5E,转Escherichia coliTG1感受态细胞,构建了单链抗体库。通过辅助噬菌体M13KO7感染后,使单链抗体展示在噬菌体衣壳蛋白pIII的N端,得到噬菌体展示的抗体库。将抗体库用于"淘洗—富集—扩增"3轮以后,得到针对脂磷酸聚糖特异性的噬菌体抗体。测序结果表明,该scFv的VH基因序列全长390 bp,编码127个氨基酸;VL基因序列全长341 bp,编码113个氨基酸。二者均符合小鼠免疫球蛋白可变区基因特征,含有4个框架区(FR)、3个抗原互补决定区(CDR)及抗体特征性的2个半胱氨酸残基。
|
| 关 键 词： | 脂磷酸聚糖 单链抗体 抗体库 |
Construction of an anti-LPG scFv library and screening for specific antibodies against LPG |
| |
| SHAN Chao,LI Lan,WANG Zong-hua.Construction of an anti-LPG scFv library and screening for specific antibodies against LPG[J].Subtropical Agriculture Research,2009,5(2). |
| |
| Authors: | SHAN Chao LI Lan WANG Zong-hua |
| |
| Abstract: | VH and VL gene fragments were first amplified from cDNA of hybridoma cell line specific to LPG,then integrated as scFv with a Linker,and cloned into phagemid vector pCANTAB-5E,and then transformed into Escherichia coli TG1 by heat-shock.To produce phage-display scFv,the helper phage M13KO7 was added to the culture when bacteria were grown up to the log-phase,in which scFv antibody was displayed in N-terminus of the bacteriophage coat protein pIII.Then the phage display antibody was applied to four rounds of "panning-enrichment-amplification" to screen for specific phage against to LPG.Our results showed that the VH gene contains 390 base pairs,encoding 127 amino acid residues.The VL gene contains 341 base pairs and encodes 113 amine acid residues.There are four FRs,three CDRs and two characteristic cysteine residues in the VH gene and the VL gene,respectively. |
| |
| Keywords: | LPG scFv antibody library |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
