His标签单克隆抗体的制备及交叉抗原的表位分析  

Preparation of monoclonal antibodies against His-tag and epitope analysis of cross antigens

作　　者：赵向绒[1] 张海祥[1] 刘杨[1] 王鑫[1] 王光华[1] 齐宗利[1] 李元[1] 胡军[1] 

机构地区：[1]陕西省人民医院中心实验室,陕西西安710068

出　　处：《细胞与分子免疫学杂志》2016年第5期683-687,692,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2014X10004002)

摘　　要：目的研究His标签对重组蛋白在疫苗、免疫和致病等方面的影响。方法制备His标签的单克隆抗体(m Ab),通过ELISA、Western blot法及免疫组织化学染色方法研究其生物学特性、免疫反应特性以及与人体组织的反应性。结果 His标签m Ab和抗原的结合与空间构型有关,多数抗His标签m Ab可以与血红蛋白结合,亦可与人体多种正常组织反应。结论带His标签的重组蛋白进行体内实验时可刺激机体产生抗His标签的抗体,该抗体可与血红蛋白及人体正常组织结合而影响重组蛋白的后期研究。Objective To explore the influence of His-tag on recombinant proteins in vaccination, immunization and pathogenesis. Methods Multiple mouse monoclonal antibodies （mAb） against His-tag were prepared. The biological and immunoreactive characteristics of these mAbs and their cross-reactivity with the normal human tissues were investigated by ELISA, Westem blotting and immunohistochemistry （IHC）, respectively. Results The binding activity of these anti-His mAbs was associated with the steric configuration of the his-tagged antigen. In addition, most of these mAbs reacted with human hemoglobin and some normal human tissues. Conclusion Anti-His antibodies could be elicited by His-tagged recombinant proteins in vivo experiments. Moreover, the functional studies of the His-tagged recombinant proteins might be affected by the reactions of anti-His6 antibodies with human hemoglobin and normal human tissues.

关 键 词：His标签 重组蛋白 单克隆抗体 空间构型 

分 类 号：Q813.2[生物学—生物工程] R392.11[医药卫生—免疫学]