人精胺氧化酶单克隆抗体的制备、鉴定及应用

发布时间：2018-06-05 09:41

本文选题：SMO + 重组蛋白　；参考：《三峡大学》2012年硕士论文

【摘要】：目的：制备鼠抗人的精胺氧化酶(Spermine oxidase, SMO)单克隆抗体，分析该抗体的效价、抗原特异性、种属特异性及在Western blot，免疫组织化学，免疫沉淀，免疫荧光，流式细胞术等分析技术中的可用性。方法：在BL21(DE3)中原核表达带6×His标签的SMO重组蛋白，并用Ni-NTA树脂亲和层析纯化。用SMO重组蛋白免疫Balb/C小鼠，通过杂交瘤技术制备抗SMO单克隆抗体，所得抗体的特异性及效价用ELISA和Western blot以及免疫组织化学等技术检测。结果：成功建立了一株稳定分泌抗人SMO的单克隆抗体杂交瘤细胞株，该单克隆抗体可用于ELISA，Western blot，免疫组织化学，免疫荧光，免疫沉淀，，流式细胞术等分析技术。结论：成功制备了高效价的SMO单克隆抗体，为SMO相关的分子、病理及抗肿瘤研究奠定了基础。
[Abstract]:Objective: to prepare mouse monoclonal antibody against human spermine oxidase (SMOA), and analyze its titer, antigen specificity, species specificity, immunocytochemistry, immunoprecipitation and immunofluorescence. Availability in analytical techniques such as flow cytometry. Methods: SMO recombinant protein with 6 脳 His tag was expressed in prokaryotic cells of BL21 DDE3 and purified by Ni-NTA resin affinity chromatography. SMO recombinant protein was used to immunize Balb/C mice. Monoclonal antibodies against SMO were prepared by hybridoma technique. The specificity and titer of the antibodies were detected by ELISA, Western blot and immunohistochemistry. Results: a hybridoma cell line with stable secretion of anti-human SMO was successfully established. The monoclonal antibody can be used for the analysis of ELISAL Western blot, immunohistochemistry, immunofluorescence, immunoprecipitation, flow cytometry and so on. Conclusion: the SMO monoclonal antibody with high titer was successfully prepared, which laid a foundation for the study of SMO related molecules, pathology and antitumor.
【学位授予单位】：三峡大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.1

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