| 抗乙型肝炎表面抗原单链抗体(ScFv)的原核表达、纯化及鉴定 |
| |
| 引用本文： | 雷清,刘鹏,郭敏,蒋琳.抗乙型肝炎表面抗原单链抗体(ScFv)的原核表达、纯化及鉴定[J].微生物学免疫学进展,2008,36(4). |
| |
| 作者姓名： | 雷清 刘鹏 郭敏 蒋琳 |
| |
| 作者单位： | 兰州生物制品研究所,兰州,730046 |
| |
| 基金项目： | 甘肃省自然科学基金
|
| |
| 摘 要： | 从抗HBsAg鼠单抗的杂交瘤细胞提取RNA,经反转录得到cDNA,进一步扩增得到鼠重链可变区基因(VH)和轻链可变区基因(VL),按VH-linker-VL的结构将VH、VL基因拼接成单链抗体(scFv)基因;经测序正确后进一步构建了表达重组体p26HBSc并在E.coli中表达,得到一约30kD的外源蛋白;纯化后经ELISA检测与HBsAg有较高的亲和力活性,为下一步的人源化改造奠定了基础。
|
| 关 键 词： | 单链抗体 连接肽 基因表达 |
Expression,purification and identification of the anti-HBsAg SeFv |
| |
| Abstract: | Obtain RNA from mouse anti-HBsAg hybridoma cell line,cDNA was got by RT,then the VH and VL gene of mouse anti-HBsAg was obtained for further amplification.According to the structure of VH-linker-VL,put VH and VL fragments together to form single chain Fv.Construct p26HBSc after sequencing and expressed in E.coli.Intrested protein was expressed about 30kD,It showed that it had higher affinity with HBsAg,so it is of potentially important for its humanization in the future. |
| |
| Keywords: | Single chain Fv(ScFv) Linker of peptide Gene expression |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
