人MUC1单克隆抗体的制备及基化MUC1抗原的表达

发布时间：2018-06-16 23:03

本文选题：MUC1 + 单克隆抗体　；参考：《吉林大学》2006年硕士论文

【摘要】： 在许多上皮来源的癌组织中,MUC1的表达上调、极性消失及糖基化程度降低使其成为多种肿瘤辅助诊断、指导治疗及判断预后的重要指标。因而大量单克隆抗体被制备用于MUC1的检测,我们决定自制抗人MUC1单克隆抗体(monoclonal antibody,McAb),建立ELISA体系检测MUC1。首先采用大肠杆菌表达的8R-MUC1-6 His为免疫原制备McAb,用间接ELISA法和免疫组化检测其效价和特异性。然以此得到的McAb无法满足制备高灵敏度及高特异性ELISA检测试剂盒的要求。为获得高效价及高特异性的McAb,我们选用Bac-to-Bac杆状病毒表达系统表达MUC1重组蛋白,此系统可以对其表达蛋白进行简单的糖基化,这一特点可使其更好的模拟肿瘤表面低糖基化的MUC1,可能成为制备McAb的理想抗原。
[Abstract]:In many epithelial cancer tissues, the up-regulated expression of MUC1, the disappearance of the polarity and the decrease of glycosylation make it an important indicator for multiple tumor assisted diagnosis, guiding treatment and judging the prognosis. Therefore, a large number of monoclonal antibodies have been prepared for the detection of MUC1, and we have determined the self-made anti human MUC1 monoclonal antibody (monoclonal antibody, McAb). The establishment of ELISA system to detect MUC1. first used 8R-MUC1-6 His expressed by Escherichia coli as immunogen to prepare McAb, using indirect ELISA and immunohistochemistry to detect its titer and specificity. However, the obtained McAb can not meet the requirements of preparing high sensitivity and high specificity ELISA detection kit. In order to obtain high efficient and highly specific McAb, I We use Bac-to-Bac baculovirus expression system to express the recombinant protein of MUC1. This system can make simple glycosylation of its expressed protein. This feature can make it better simulate the low glycosylated MUC1 of the tumor surface, and may be an ideal antigen for the preparation of McAb.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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