摘要
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 the hybridoma cell lines excreting monoclonal antibodies against granulocytecolony stimulating factor(GCSF).Methods:The mouse SP2/0 myeloma cells and splenocytes from Balb/c mice immunized with GCSF were fused by application of PEG1500.The monoclonal antibodies produced by these hybridoma cells were identified by enzymelinked immunosorbent assay(ELISA).Resu lts:Five hybridoma cell lines secreting antiGCSF monoclonal antibodies were acquired.Monoclonal antibody titers in the supernants of the incubation media rose to 1∶103 and in the ascites of the mice inoculated the hybridoma cells,1∶107.Chromosome of hybridoma cells had normal morphology and its number was 83～92.These hybridoma cell lines fairly reserved the antibodyproducing capacity after stably growing in vitro for four to six months and through frozing and thawing.Conclusion:We have successfully established hybridoma cell lines with high titers and stably producing antiGCSF monoclonal antibodies by PEG fusing techniques .It will provide conditions for developing ELISA kit of monoclonal antibodies directed to GCSF.
出处
《滨州医学院学报》
1998年第4期315-317,共3页
Journal of Binzhou Medical University
关键词
单克隆抗体
杂交瘤细胞
G-CSF
granulocytecolony stimulating factor
monoclonal antibodies
hybridoma cell lines
