用双向电泳及蛋白质印迹技术筛选乳腺癌相关抗原

目的：应用双向电泳及免疫印迹技术分析乳腺癌MCF-7细胞总蛋白,寻找乳腺癌相关抗原。方法：提取乳腺癌MCF-7细胞总蛋白,双向电泳技术（2-DE）对总蛋白进行分离后转膜,将健康人血清（对照组）和乳腺癌患者血清（实验组）作为一抗进行蛋白质印迹（Western blotting）分析,寻找杂交蛋白点的差异。结果：MCF-7总蛋白双向电泳考马斯亮蓝染色图谱共检测到248个蛋白质斑点,Western blotting显示实验组抗原抗体反应点23个,对照组为13个,找到10个差异蛋白点。结论：乳腺癌患者血清中的抗体表达水平有改变,这10个差异蛋白点有可能是乳腺癌相关抗原。

Screening of Human Breast Cancer Associated Antigens with Two Dimensional Electrophoresis and Western Blot

Objective： To analyze total protein of mammary cancer cells（MCF-7 cells） with two dimensional electrophoresis and western blot,and to search for human breast cancer associated antigens.Methods： Total proteins in MCF-7 cells that cultured in vitro were extracted and separated by two-dimensional electrophoresis（2-DE）.The separated protein bands were then transfered to polyvinylidene fluoride（PVDF） membrane,and incubated with serums of healthy people（group C） or patients（group E） as the source of primary antibody for western blotting and the resulted spots were used for protein analysis.Results： Two hundred and forty-eight spots of total protein in MCF-7 cells were found by 2-DE,and the numbers of antigen reaction spots showed by western blotting were 23 in group E,and 13 in group C.There were 10 contrast protein spots.Conclusions： Antibody expression in patient serum has changed.The 10 differently expressed protein spots might be breast cancer associated antigens.