抗人hER单克隆抗体的制备及其应用

目的:制备抗人hER单克隆抗体。方法:用原核表达的人雌激素受体(hER)重组蛋白为抗原,免疫BalB/c小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤SP2/0细胞,进行细胞融合。结果:通过ELISA和免疫组织化学法筛选,有限稀释克隆化,得到一株分泌抗hER单克隆抗体的杂交瘤细胞株(2B8/1C5/2C3),经鉴定2B8/1C5/2C3分泌的单克隆抗体为IgG2a,利用Western Blot分析,显示在23kU处针对hER重组蛋白位置有一特异性条带;利用乳腺癌组织进行的免疫组织化学染色结果显示该株细胞产生的单克隆抗体与细胞核呈强阳性反应。结论:该单克隆抗体适于乳腺癌组织hER的免疫组化检测。

Preparation of Monoclonal Antibody against Recombinant Protein of Human Estrogen Receptor

Objective: To prepare the monoclonal antibodies (McAbs) of human estr og en(hER). Methods: McAbs were preparated against hER by th e cell fusion of immunized BalB/c mouse spleen cell and mouse myeloma cell SP2/0. The antigen hER was recombinant protein expressed by prokaryotic expression syst em. Results: After several times of limited dilution, one clone (2B8/1C5/2C3) of hybridoma cells was obtained by ELISA screening and immunocyto chemistry. The isotype of the pure 2B8/1C5/2C3 McAb was IgG2a. With Western blot analysis, a single strong immunostained blot appeared at molecular weight of 23 kD. In immmunohistochemistry staining, there appeared strong cell nuclear stain ing on paraffinembeded breast cancer sections. Conclusion: Th e McAb was suitable for detection of hER of breast cancer by immunocytochemistry .