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目的制备并鉴定抗人前列腺干细胞抗原(PSCA)的单克隆抗体。方法以纯化的PSCA为抗原免疫Balb/c小鼠,经细胞融合、克隆化培养建立抗人PSCA的杂交瘤细胞株,采用免疫组化、Westernblot等方法鉴定其特异性。结果利用小鼠杂交瘤技术,建立了1株分泌抗PSCA抗体的杂交瘤细胞系D3,经鉴定抗体类别为IgG1,能特异性识别PSCA,与其他因子无交叉反应。PSCA在5例正常前列腺组织、10例良性前列腺增生组织、38例前列腺癌组织中有不同程度的阳性染色,95%(38/40例)的前列腺癌PSCA染色呈阳性、强阳性染色,50%~100%的肿瘤细胞着色,与良性前列腺增生及正常前列腺组织相比,其染色强度及范围的差异有统计学意义(P<0.01)。结论成功制备了组织特异性的抗人PSCA单克隆抗体,为前列腺癌的诊断和免疫治疗提供了新的手段。
Objective To prepare and identify monoclonal antibodies against human prostate stem cell antigen (PSCA). Methods Balb / c mice were immunized with purified PSCA as antigen. After hybridization and cloning, the anti-human PSCA hybridoma cell lines were established and characterized by immunohistochemistry and Western blot. Results A hybridoma cell line D3 secreting anti-PSCA antibody was established by using the mouse hybridoma technology. The antibody was identified as IgG1, which recognized PSCA specifically and did not cross-react with other factors. PSCA had positive staining in 5 cases of normal prostate, 10 cases of benign prostatic hyperplasia and 38 cases of prostate cancer, PSCA staining in 95% (38/40) of prostate cancer was positive, strong positive staining, 50% ~ 100% of the tumor cell staining, compared with benign prostatic hyperplasia and normal prostate tissue, the staining intensity and range of difference was statistically significant (P <0.01). Conclusion Tissue-specific anti-human PSCA monoclonal antibody was successfully prepared and provided a new method for the diagnosis and immunotherapy of prostate cancer.