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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。
BALB/c mice were immunized with activated human B cell line 3D5 cells, and mouse spleen cells were fused with SP2/0 cells. After fusion, the cells were grown on methylcellulose semi-solid medium. The 0.5% formaldehyde-treated 3D5 cells and CEM cells were coated with the enzyme-labeled plates as target cells, and 290 hybridoma cell clones were screened using a cell-enzyme-linked immunosorbent assay (CELISA) assay, resulting in 33 positive reactions with 3D5 cells. CEM cell-negative clones. After indirect immunofluorescence staining, the 33 positive clones obtained above were retested by flow cytometry. Results showed that 27% of 3D5-positive CEM-negative individuals accounted for 81.82%, indicating that CELISA is a rough screening anti-human B cell differentiation antigen. Easy and effective method.