摘要
目的 :制备分泌抗乙型肝炎病毒末端蛋白mAb杂交瘤并研究杂交瘤细胞及所产生抗体的特性 .方法 :用经原核表达的乙型肝炎病毒末端蛋白 (HBV TP)免疫BALB/c小鼠 ,取其脾细胞与骨髓瘤细胞SP2 /0融合 ,经筛选和克隆化建立杂交瘤细胞系 ,然后用ELISA法筛选出阳性克隆 .以免疫组化ABC方法研究杂交瘤细胞分泌的mAb特性 .结果 :共筛选出 3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 1 0 0d连续培养 ,mAb分泌稳定 ,特异性强 ,腹水抗体效价免疫组化ABC法 1∶1 0 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 (80 % ) ,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 3株抗乙型肝炎病毒末端蛋白杂交瘤细胞分泌的mAb具有特异亲合性 。
AIM: To establish anti HBV Terminal Protein (TP) antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. METHODS: The BALB/c mice were immunized with HBV Terminal 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 plated and the antibodies against HBV RT from the positive clones were further identified with the immunobistochemistry method. RESULTS: There were three hybiridoma cell lines established, both of them secreted high quality mAbs steadily and all cells had the characters of hybridoma. It was found that more than 80 percent specimens of HBV related liver diseases showed positive staining, while there was no positive staining in normal hepatocytes. CONCLUSION: Successful generation of high quality hybridomas secreting anti HBV TP monoclonal antibodies will facilitate both early diagnosis and treatment of HBV infection.
出处
《第四军医大学学报》
北大核心
2003年第11期1000-1001,共2页
Journal of the Fourth Military Medical University
关键词
HBV
末端蛋白
杂交瘤
单克隆抗体
免疫组化
HBV
Terminal Protein
hybridoma
monoclonal antibody
immunohistochemistry
