抗PSMA膜外段单克隆抗体的制备及其初步应用

Preparation and Preliminary Utilization of Monoclonal Antibodies Specifically Against Extracellular Peptide Fragment of Prostate Specific Membrane Antigen

目的 制备抗前列腺特异性膜抗原(PSMA)膜外段的单克隆抗体。方法 原核表达含有PSMA膜外段多肽的融合蛋白,免疫雌性Balb/c小鼠后取效价较高的小鼠行小鼠脾细胞与SP2/0细胞融合,以酶联免疫吸附实验(ELISA)、免疫细胞化学、免疫组织化学法筛选、鉴定分泌特异性抗PSMA膜外段的单克隆杂交瘤细胞株。结果 经过ELISA初筛得到55个阳性杂交瘤细胞株,其中8株经免疫组化证实为可用于前列腺癌石蜡包埋组织切片的免疫组化染色。结论 获得了8株可持续分泌抗PSMA膜外段单克隆抗体的杂交瘤细胞株,产生的单抗以后可用于前列腺癌的诊断,并为前列腺癌的免疫治疗奠定了基础。

Purpose: To prepare monoclonal antibodies specifically against extracellular peptide fragment of prostate specific membrane antigen (PSMA). Methods: The extracellular fragment of PSMA protein expressed by a prokaryotic expressing system was used to immunize female Balb/c mice, splenocytes from immunized Balb/c mice were fused with SP2/0 cells. The hybridoma cells were primarily screened by enzyme-linked immunoabsorbent assay (ELISA), and the positive clones were further identified by immunocytochemistry, immunohistochemistry techniques. Results: Fifty-five clones were primarily selected as positive by ELISA, and finally eight clones were identified as positive by immnohistochemistry and can be used in the slides of formalin-fixed, paraffin-embedded tissue of prostate carcinoma. Conclusions: Eight hybridoma cell lines secreting monoclonal antibodies specifically against the extracellular peptide fragment of PSMA were prepared, which may be used in the diagnosis of prostate carcinoma, and as a tool for the immnotherapy of prostate carcinoma.