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目的：建立鼠抗粒细胞集落刺激因子（Ｇ－ＣＳＦ）单克隆抗体的杂交瘤细胞系。方法：将ＳＰ２／０小鼠骨髓瘤细胞和用Ｇ－ＣＳＦ免疫的Ｂａｌｂ／ｃ小鼠脾细胞在ＰＥＧ－１５００作用下进行融合；用ＥＬＩＳＡ进行克隆化筛选并对杂交瘤细胞分泌的单抗滴度进行检测。结果：获得了５株分泌抗Ｇ－ＣＳＦ单克隆抗体的杂交瘤细胞株；细胞培养上清液的单抗滴度可达１∶１０３，杂交瘤细胞注射同系小鼠制备的腹水单抗滴度达１∶１０７；杂交瘤细胞染色体数基本上是８３～９２条，形态正常，在体外连续传代４～６个月，经冻存复苏后仍能正常分泌特异性抗体。结论：利用ＰＥＧ融合技术成功地建立了稳定产生高效价的抗Ｇ－ＣＳＦ单抗的杂交瘤细胞系，为开发研制抗Ｇ－ＣＳＦ单抗的ＥＬＩＳＡ试剂提供了条件。
Objective: To establish a hybridoma cell line with murine anti-granulocyte colony-stimulating factor (G-CSF) monoclonal antibody. METHODS: SP2/0 mouse myeloma cells and Balb/c mouse spleen cells immunized with G-CSF were fused with PEG-1500; clones were screened by ELISA and the monoclonal antibody secreted by hybridoma cells was dripping. Degree of detection. RESULTS: Five hybridoma cell lines secreting anti-G-CSF monoclonal antibodies were obtained; the titer of the monoclonal antibody in the cell culture supernatant was 1:103. The ascites monoclonal antibody titer prepared by the hybridoma cells injected into syngeneic mice was obtained. Up to 1:107; the hybridoma cell chromosome number is basically 83 ~ 92, the normal morphology, continuous passage in vitro 4 to 6 months, after cryopreservation can normally secrete specific antibodies. Conclusion: The hybridoma cell line stably producing high titer anti-G-CSF monoclonal antibody was successfully established by PEG fusion technology, which provided the conditions for the development of ELISA reagents for anti-G-CSF monoclonal antibody.