【摘要】 目的 制备抗肝癌特异性谷氨酰转肽酶(GGT)单克隆抗体及研究此单克隆抗体的临床应用。 方法 以纯品GGT-Ⅱ免疫BALB/C小鼠,取免疫鼠脾细胞与SP2/0骨髓瘤细胞融合以制备能产生抗GGT-Ⅱ单克隆抗体的杂交瘤细胞株;以此抗GGT-Ⅱ单克隆抗体采用竞争抑制性ELISA检测人血清GGT-Ⅱ。 结果对融合细胞阳性孔反复克隆后获得1株稳定分泌抗GGT-Ⅱ单克隆抗体的小鼠杂交瘤细胞系:2G4F6B2,此单克降抗体为IgG1类,与GGT-I无交叉反应。以此单克隆抗体测定人血清GGT-II的结果与聚丙酰胺梯度电泳法相符。 结论 研究制备的抗GGT-Ⅱ单克隆抗体特异性高,可应用于临床检测人血清中GGT-Ⅱ。更多还原

【Abstract】 Objective To prepare a monoclonal antibody against hepatoma-specific gamma-glutamyltransferase (GGT-Ⅱ) and study it’s application. Methods Two Bal B/C mice were immunized with pure GGT- Ⅱ , then their spleen cells were separated and fused to SP 2/0 myeloma cells so as to make hybridoma cell strain which could yield monoclonal antibody against GGT- Ⅱ. And it’s effect of binding GGT- Ⅱ was detected by competitive inhibitory enzyme linked-immunosorbance assay (ELISA). Results A mouse hybridoma cell strain which could steadily secrete the monoclonal antibody against GGT- Ⅱ was obtained and named 2G4F6B2. This monoclonal antibody belonged to IgG1 subclass and was specific to GGT- Ⅱ , without cross-reaction to GGT- I . The result of detecting human serum GGT-Ⅱ by ELISA with the monoclonal antibody accorded with that by polyacrylamide gradient electrophoresis. Conclusion The monoclonal antibody against GGT- Ⅱ prepared in this study has high specificity and can be applied in clinic to detect human serum GGT- Ⅱ.更多还原