大鼠抗小鼠重组膜联蛋白A2单克隆抗体的制备与鉴定  

Preparation and Identification of Rat mAb Against Recombinant Murine Annexin II

作　　者：江淼[1] 沈飞[1] 包红雨[1] 张丽意[1] 谢丽倩[1] 赵益明[1] 阮长耿[1] 

机构地区：[1]苏州大学附属第一医院江苏省血液研究所卫生部血栓与止血重点实验室,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2011年第6期879-882,892,共5页Suzhou University Journal of Medical Science

基　　金：美国血液学会发展中国家研究培训基金资助项目(ASH20060108)

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。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.

关 键 词：单克隆抗体 杂交瘤细胞系 膜联蛋白A2 

分 类 号：R392-22[医药卫生—免疫学]