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用细胞酶联免疫吸附法筛选分泌抗人B细胞分化抗原单抗的杂交瘤

Screening of Hybridoma Secreting McAb to Human B Celldifferentiation Antigen(s)Using Cellular Enzyme-linked Immunoabsorbentassay

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【作者】 郭彩云石伟王汛朱立平

【Author】 Guo Caiyun Shi Wei Wang Xun Zhu Liping (Institute of Basic medical Scienecs. Chinese Academy of Medical Scienecs,Beijing 100005)

【机构】 中国医学科学院基础医学研究所

【摘要】 用活化的人B细胞株3D5细胞免疫BALB/c小鼠,取小鼠脾脏细胞与SP2/0细胞融合。融合后细胞置甲基纤维素半固体培养基生长。以0.5%甲醛处理的3D5细胞和CEM细胞包被酶标板作为靶细胞,用细胞酶联免疫吸附(CELISA)试验筛选290个杂交瘤细胞克隆,得到33个与3D5细胞反应阳性而与CEM细胞反应阴性的克隆。再经间接免疫荧光染色后,用流式细胞仪复测以上所得33个阳性克隆,结果3D5阳性CEM阴性者27例,占81.82%,表明CELISA法是一个粗筛抗人B细胞分化抗原的简便有效方法。

【Abstract】 plenocytes from BALB/C mouse immunized with 3D5 cell, an activated human B cell line,were fused with SP2/0 cells. The fused cells grew in a methyl cellulose semi-solid medium. 290hybridoma clones were obtained. By using 0.5% methynol-treated 3D5 cells and CEM cells,ahuman T cell line, coated on ELISA plates as targets for screening, 33 of 290 clones were foundto secrete McAbs which reacted with 3D5 cells, but not with CEM cells. To confirm the results,the McAbs in the supernatants from the 33 clones were detected for thair reactivity to 3D5 cells orCEM cells upon a flowcytometerbased on indirect immunofluorescent staining. 27 of 33 clones(81. 82%)still showed positive reactions to 3D5 cells and negative reactions to CEM cells indicat-ing that CELISA is a primary approach for screening McAbs to human B cell differentiation anti-gens with convenience and reliability.

【关键词】 3D5细胞细胞酶联免疫吸附试验单克隆抗体
【Key words】 D5 cell CELISA McAb
  • 【文献出处】 基础医学与临床 ,BASIC MEDICAL SCIENCES AND CLINICS , 编辑部邮箱 ,1995年02期
  • 【分类号】R730.3
  • 【被引频次】3
  • 【下载频次】52
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