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

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。

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.