【摘要】 目的建立能够分泌特异性抗小鼠膜联蛋白A2(AnxA2)的大鼠-小鼠融合单克隆抗体杂交瘤细胞系。方法利用重组小鼠AnxA2蛋白作为抗原免疫Wistar大鼠,通过细胞融合和杂交瘤筛选技术建立特异分泌抗小鼠AnxA2蛋白的大鼠-小鼠融合单抗杂交瘤细胞株。采用酶联免疫吸附方法确定单克隆抗体亚型,免疫印迹和流式细胞术验证所筛选的单克隆抗体的特异性。结果通过对30多个分泌抗体的融合细胞克隆的筛选,获得一株分泌特异性抗小鼠AnxA2的大鼠-小鼠融合单克隆抗体杂交瘤细胞系SZ-158,连续培养传代后染色体分析证实,Wistar大鼠脾细胞与小鼠SP2/0细胞稳定融合;免疫印迹和流式细胞术显示该单克隆抗体可以特异识别小鼠AnxA2重组蛋白。结论该研究所获得的大鼠-小鼠融合单克隆抗体特异性针对小鼠AnxA2蛋白,可用于AnxA2转基因小鼠中AnxA2表达的检测,为深入研究AnxA2的功能提供了物质保障。更多还原

【Abstract】 Objective To establish a rat-murine fusion hybridoma cell line secreting monoclonal antibody against murine annexinⅡ(Anx A2).Methods Recombinant murine annexinⅡprotein was used as antigen,and rat-murine cell fusion and screening technology was used to establish the hybridoma cell line.The subtype of the monoclonal antibody was detected by enzyme-linked immunosorbent assay. And the specificity of the monoclonal antibody against murine endoglin was tested by Western blot and flowcytomery.Results More than 30 antibody-producing clones were screened.One clone,named as SZ-158,was proved to have the capability of stably secreting monoclonal antibody against murine annexinⅡ.The subtype of mAb SZ-158 was IgGl,and the light chain was kappa.It was demonstrated that mAb SZ-158 specifically recognized the recombinant murine annexinⅡby Western blot and flowcytomery analysis. Conclusion mAb SZ-158 can be used as a powerful tool for function studies in annexinⅡgene transfer murine models.更多还原