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以往的研究发现 6 3 6 4 %的胰腺癌患者具有针对相对分子质量初步鉴定为 4 70 0 0的靶抗原的阳性共带 ,该条带仅在1 9%的正常人中查及 ,本研究旨在纯化并鉴定该靶抗原 ,以深入认识该独特的自身免疫现象在胰腺癌中的意义 ,探索靶抗原的生物学功能以及在临床诊断中可能应用价值。提取人喉癌上皮细胞 (Hep 2 )蛋白抗原 ,进行双向电泳 ,选取 8份以往在单向蛋白质印迹 (SDS PAGE )中 ,相对分子质量约为 4 70 0 0处有明显抗原抗体反应的胰腺癌患者血清进行免疫印迹分析。比较双向电泳凝胶图像、丽春红染色之蛋白质转印硝酸纤维素滤膜图像以及血清反应后滤膜图像 ,确定靶抗原在凝胶上的斑点 ,判断其等电点。切取胶上蛋白质斑点经胰蛋白酶消化后通过HPLC MS联用分析获得串级质谱数据 ;利用互联网搜索工具MASCOT分析处理数据并进行数据库查询 ,以鉴定靶抗原。结果表明 ,KIAA0 111的基因预测产物是一种新鉴定的胰腺癌相关性自身抗体靶抗原 ,有希望成为新的胰腺癌标志物
Previous studies found that 63.64% of pancreatic cancer patients had a positive conferrence against a target antigen preliminarily identified as 4 70 0 relative molecular weight, which was only found in 19% of normal individuals. This study To purify and identify the target antigen in order to gain insight into the unique significance of autoimmune phenomena in pancreatic cancer, to explore the biological function of target antigen and its possible value in clinical diagnosis. The antigen of human laryngeal carcinoma epithelial cells (Hep 2) was extracted and subjected to two-dimensional electrophoresis. Eight samples of pancreatic cancer that had a significant antigen-antibody reaction at a molecular weight of about 4 70 0 0 in a single-pass Western blot (SDS PAGE) Patient sera were subjected to immunoblot analysis. Comparison of two-dimensional electrophoresis gel images, Ponceau-stained image of the protein-transferred nitrocellulose filter and the serum-reacted filter image to determine the target antigen on the gel spots to determine the isoelectric point. The tandem mass spectrometry data was obtained by digesting the protein spots on the gel by trypsinization and analyzing by MS-MS. The data were searched by MASCOT, an Internet search tool, and the database was searched to identify target antigens. The results show that the predicted gene of KIAA0 111 is a newly identified target antigen of pancreatic cancer-associated autoantibodies, promising to be a new pancreatic cancer marker