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目的:研制可识别天然CD100分子的单克隆抗体(mAb)。方法:采用真核表达的CD100分子免疫BALB/c小鼠,以间接ELISA法筛选阳性杂交瘤,并分别用人急性T淋巴细胞性白血病细胞系MOLT-4、猿T淋巴细胞系MLA-144及猴EB病毒转化的B淋巴细胞系(BLCL),进行间接免疫荧光染色和流式细胞术鉴定mAb的特异性。结果:共获得7株可稳定分泌 mAb的杂交瘤细胞株。所有mAb均可识别人和猴细胞系的膜表面天然CD100分子,其中6株可识别猿细胞系膜表面的天然CD100分子。结论:成功地制备了7株抗CD100分子的特异性mAb,为研究人和猿、猴CD100分子的结构与功能提供了新的手段。
Objective: To develop a monoclonal antibody (mAb) that recognizes natural CD100 molecules. Methods: BALB / c mice were immunized with eukaryotic expression vector CD100 and the positive hybridomas were screened by indirect ELISA. The positive clones were selected by using human acute lymphoblastic leukemia cell line MOLT-4, ape T lymphocyte line MLA-144 and monkey Epstein-Barr virus-transformed B lymphocyte cell line (BLCL) was subjected to indirect immunofluorescence staining and flow cytometry to identify the specificity of the mAb. Results: Seven hybridoma cell lines that can stably secrete mAb were obtained. All mAbs recognize native CD100 molecules on the membrane surface of human and monkey cell lines, of which 6 are native CD100 molecules that recognize the surface of simian cell membranes. Conclusion: Seven anti-CD100 specific mAbs were successfully prepared, which provided a new method for studying the structure and function of human and apes, monkey CD100.