基础医学与临床 ›› 2010, Vol. 30 ›› Issue (9): 952-955.

• 研究论文 • 上一篇    下一篇

表达膜抗AchR抗体的杂交瘤细胞株的建立

赵青 尹娜 张伟   

  1. 中国医学科学院基础医学研究所北京协和医学院基础学院 中国医学科学院基础医学研究所
  • 收稿日期:2010-02-24 修回日期:2010-03-29 出版日期:2010-09-05 发布日期:2010-09-05
  • 通讯作者: 张伟

Establishment of hybridoma cell strains expressing membrane- specific antibody against AchR

Qing ZHAO, Na YIN, Wei ZHANG   

  1. Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences,School of Basic Medicine, Peking Union Medical College IBMS, CAMS & PUMC
  • Received:2010-02-24 Revised:2010-03-29 Online:2010-09-05 Published:2010-09-05
  • Contact: Wei ZHANG

摘要: 目的 建立表达膜特异性自身抗体的细胞株。方法 用DNA重组技术表达乙酰胆碱受体 亚基,并用其免疫小鼠。应用杂交瘤技术,建立小鼠脾细胞融合的杂交瘤细胞株。通过流式细胞术分析杂交瘤细胞膜抗体和膜免疫球蛋白的表达,SDS聚丙烯酰胺凝胶电泳分析膜抗体和分泌型抗体分子量的差别,用免疫沉淀和Western blot等方法鉴定膜蛋白和分泌蛋白。结果 杂交瘤细胞既可以分泌抗体又可以表达膜抗体。结论 表达膜抗AchR的杂交瘤细胞类似于重症肌无力中表达膜自身抗体的B细胞,可进一步作为靶细胞研究该疾病。

关键词: 杂交瘤细胞, 膜抗体, 重症肌无力

Abstract: Objective To establish cell strains that express membrane-specific anti-AchR autoantibodies. Methods The extracellular reagion of AchR subunit was expressed and used as an immunogen to immunize mice. Using hybridoma technology to get anti-AchR antibody cell strains. Flow cytometry analysis was performed to detect membrane specific antibody and membrane Ig expression. SDS-PAGE was performed to compare molecular weight of membrane and secreted antibodies. Immunoprecipitation and Western blot were performed to charicterize membrane and secretary antibodies. Results Hybridoma cells could not only secrete antibodies, but also expresse membrane antibodies. Conclusion Membrane anti-AchR antibody expressing hybridoma cells were similar as membrane autoantibody expression B cells in Myasthernia Gravis and may be used as target cells for studying this disease.

Key words: hybridoma cell, membrane antibody, Myasthernia Gravis

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