论文部分内容阅读
为制备出具较高特异性抗肝癌核基质蛋白单克隆抗体，以用于肝癌的基础和临床的研究。首次免疫用肝癌细胞纯核，用高盐抽提法提取的肝癌细胞核基质蛋白加强免疫Ｂａｌｂ／ｃ小鼠，取其脾细胞与ＳＰ２／０骨髓瘤细胞融合，ＥＬＩＳＡ法同时测定培养上清对肝癌核基质蛋白及正常肝细胞核基质蛋白的免疫反应，ＬＳＡＢ法进行单抗的鉴定。结果：建立了一株能稳定分泌抗肝癌核基质蛋白单抗的杂交瘤细胞株２Ｇ８，染色体数目为８６－９２，Ｉｇ亚类鉴定为ＩｇＭ，它与ＨＢｓＡｇ、ＨＢｅＡｇ等ＨＢＶ相关抗原无交叉反应，与ＡＦＰ亦无阳性反应，ＬＳＡＢ法证实单抗定位于肝癌细胞核上，与正常肝及胎肝细胞没有交叉反应，与４种癌症细胞和３种正常细胞的反应发现仅食管癌细胞核上有较弱的阳性反应。结论：获得一株较特异的抗肝癌核基质蛋白单抗，可望进一步用于肝癌的诊断治疗及肝癌发生发展的研究。
To produce a higher specificity of anti-liver cancer nuclear matrix protein monoclonal antibody, for the basic and clinical research of liver cancer. Balb / c mice were immunized with the pure nuclei of hepatocellular carcinoma cells for the first time. The Balb / c mice were immunized with nuclear matrix proteins extracted from the liver cancer cells by high salt extraction. The spleen cells were fused with SP2 / 0 myeloma cells. Nuclear matrix protein and normal liver cell nuclear matrix protein immune response, LSAB method for the identification of monoclonal antibody. Results: A hybridoma cell line 2G8 was established which can stably secrete anti-hepatoma nuclear matrix protein monoclonal antibody. The number of chromosomes was 86-92. The Ig subclass was identified as IgM. It had no cross-reaction with HBV-related antigens such as HBsAg and HBeAg. There was no positive reaction with AFP. The LSAB method confirmed that the McAb was located on the nucleus of hepatoma cells and had no cross-reaction with normal liver and fetal hepatocytes. The reaction with 4 kinds of cancer cells and 3 kinds of normal cells found only weak nuclei in esophageal cancer cells Positive reaction. Conclusion: Obtaining a specific anti-hepatoma nuclear matrix protein monoclonal antibody is expected to be further used in the diagnosis and treatment of liver cancer and the development of liver cancer.