| 抗微囊藻毒素单链抗体基因的构建、表达及初步鉴定 |
| |
| 引用本文： | 宋丽敏,张维,陈明,林敏,潘家荣.抗微囊藻毒素单链抗体基因的构建、表达及初步鉴定[J].高技术通讯,2010,20(3). |
| |
| 作者姓名： | 宋丽敏 张维 陈明 林敏 潘家荣 |
| |
| 作者单位： | 1. 中国农业科学院农产品加工研究所,北京,100193
2. 中国农业科学院生物技术研究所,北京,100081 |
| |
| 摘 要： | 利用RT-PCR方法从抗微囊藻毒素-LR(MC-LR)单克隆抗体的杂交瘤细胞中扩增出抗体的V_H和V_L基因,构建了抗MC-LR分子的单链抗体(scFv)基因。SDS-PAGE和Western blot分析结果显示,该单链抗体基因在大肠杆菌Origami 2中特异性表达出分子量约为30kD的融合蛋白。通过Ni-NTA金属亲和层析法对可溶性表达产物进行纯化,获得的目的蛋白浓度为0.115mg/mL。ELISA反应结果表明该单链抗体能与MC-LR特异性结合。此研究为制备多价高亲和力抗MC-LR抗体奠定了基础。
|
| 关 键 词： | 微囊藻毒素(MC) 单链抗体 原核表达 |
Construction, expression and identification of scFv against microcystin-LR |
| |
| Song Limin,Zhang Wei,Chen Ming,Lin Min,Pan Jiarong.Construction, expression and identification of scFv against microcystin-LR[J].High Technology Letters,2010,20(3). |
| |
| Authors: | Song Limin Zhang Wei Chen Ming Lin Min Pan Jiarong |
| |
| Abstract: | The V_H and V_L genes were amplified by RT-PCR from hybridoma cell secreting anti-microcystin-LR monoclonal antibody, and the scFv gene was spliced by sequence overlap extension PCR with V_H and V_L SDS-PAGE and Western blot analysis confirmed the expression of scFv with the molecular weight of about 30kD. The recombinant protein was purified by metal affinity chromatography using Ni-NTA, and the concentration of purified scFv was 0.115mg/mL. The ELISA assay revealed that the scFv protein could bind specifically to MC-LR. The results provide a basis for the application of genetic engineering methods in the preparation of multivalent antibodies with high affinity. |
| |
| Keywords: | ELISA microcystins (MC) single chain variable fragment prokaryotic expression ELISA |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
