摘要
目的 建立抗乙型肝炎病毒逆转录酶单克隆抗体杂交瘤并研究杂交瘤细胞及抗体特性 .方法 用经原核表达的乙型肝炎病毒逆转录酶蛋白 (HBV- RT)免疫 BAL B/ c小鼠 ,取其脾细胞与骨髓瘤细胞 Sp2 / 0融合 ,经筛选和克隆化制备杂交瘤细胞系 ,然后用 EL ISA法筛选出阳性克隆 .最后用免疫组化 ABC方法研究杂交瘤细胞分泌的单克隆抗体特性 .结果 2株能持续、稳定分泌抗体的杂交瘤细胞系 ,其杂交瘤细胞经过 10 0 d连续培养 ,单克隆抗体分泌稳定、特异性强、腹水抗体效价免疫组化 ABC法 1∶ 10 0 0 .免疫组化检测显示其相应抗原在肝炎及肝硬变组织中呈高表达 (73% ) ,正常肝组织中未见阳性反应 ;其抗原主要分布于细胞质内 .结论 此2株抗乙型肝炎病毒逆转录酶杂交瘤细胞分泌的单克隆抗体具有特异亲合性 ,它们的成功制备为研究
AIM To establish anti HBV reverse transcriptase antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. METHODS The BALB/c mice were immunized with HBV reverse transcriptase protein (HBV RT) and the immunized mouse spleen cells were fused with SP2/0 cells to raise hybridomas after three times of inoculation. The positive clones of hybridomas were screened by ELISA methods using RT protein coated 96 well plate and the antibodies against HBV RT from the positive clones were further identified with the immunohistochemistry method. RESULTS There were two hybridoma cell lines established and both of them secreted high quality mAbs steadily and all cells had the characters of hybridoma. It was found that more than 73 percent specimens of HBV related liver diseases showed positive staining, while there was no positive staining in normal hepatocytes. CONCLUSION The successful generation of high quality hybridomas secreting anti HBV RT monoclonal antibodies will facilitate both early diagnosis and treatment of HBV infection.
出处
《第四军医大学学报》
2000年第4期446-448,共3页
Journal of the Fourth Military Medical University
