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目的制备抗人胰腺癌细胞单克隆抗体(McAb),并对其特性进行鉴定。方法以胰腺癌细胞系Patu8988免疫Balb／ c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2／0进行融合,经选择培养,筛选及克隆化后,建立杂交瘤细胞系。以ELISA flow cytometry等方法研究抗体特性。以MTT法研究细胞毒作用。结果得到一株能够稳定分泌特异性抗体的杂交瘤细胞系 A7。用琼脂糖双扩散鉴定其为IgG1亚类,ELISA测定其腹水效价为:1:16000。流式细胞仪检测表明:该抗体与Patu8988 细胞呈阳性反应(>90％),与肝癌、结肠癌、胃癌、肺癌、乳腺癌呈阴性反应(<5％)。MTT法示该抗体对胰腺癌细胞的杀伤率明显高于对照组。结论 McAbA7对胰腺癌的诊断和治疗可能具有一定的临床意义。
Objective To prepare anti-human pancreatic cancer monoclonal antibody (McAb) and identify its characteristics. Methods Balb / c mice were immunized with pancreatic cancer cell line Patu8988. The spleen cells were fused with mouse myeloma cells SP2 / 0. After selective culture, screening and cloning, the hybridoma cell lines were established. Antibody characteristics were studied by ELISA flow cytometry. MTT assay cytotoxicity. As a result, a hybridoma cell line A7 capable of stably secreting specific antibodies was obtained. It was identified as IgG1 subclass by double diffusion of agarose. The ascites titer was 1: 16000 by ELISA. Flow cytometry showed that the antibody was positive (> 90%) to Patu8988 cells and negative to liver cancer, colon cancer, gastric cancer, lung cancer and breast cancer (<5%). MTT method showed that the antibody killing rate of pancreatic cancer cells was significantly higher than the control group. Conclusion McAbA7 may be of clinical significance in the diagnosis and treatment of pancreatic cancer.