Kimura H, Kudo T, Nishihara S, Iwasaki H, Shinya N, Watanabe R, Honda H, Takemura F, Narimatsu H
Institute of Life Science, Soka University, Tokyo, Japan.
Glycoconj J. 1995 Dec;12(6):802-12. doi: 10.1007/BF00731242.
We prepared a mouse monoclonal antibody, FTA1-16, that specifically recognizes human alpha(1,3/1,4)fucosyltransferase without crossreactivity to any other members of the alpha(1,3)fucosyltransferase family. The specificity was confirmed by both immunofluorescense staining of native antigens in the Golgi apparatus and Western blotting analysis, using stable transformant cells transfected with each gene of the alpha(1,3)fucosyltransferase family. Western blotting analysis on a series of human tumour cell lines from various tissues revealed that some epithelial cancer cell lines from digestive organs expressed an amount of alpha(1,3/1,4)fucosyltransferase in good correlation with expression of sialyl Lewis a antigen. Immunohistochemical staining by FTA1-16 on colon cancer tissues revealed enhanced expression of the enzyme in cancer cells in comparison to normal cells. Finally, the antigenic epitope recognized by FTA1-16 was determined using truncated recombinant peptides which were expressed in E. coli. A minimal length determined was a fragment, amino acid positions 132-153, of the alpha(1,3/1,4)fucosyltransferase.
我们制备了一种小鼠单克隆抗体FTA1-16,它能特异性识别人类α(1,3/1,4)岩藻糖基转移酶,且与α(1,3)岩藻糖基转移酶家族的其他成员无交叉反应。通过使用转染了α(1,3)岩藻糖基转移酶家族各基因的稳定转化细胞,对高尔基体中的天然抗原进行免疫荧光染色和蛋白质印迹分析,证实了其特异性。对一系列来自不同组织的人类肿瘤细胞系进行蛋白质印迹分析发现,一些来自消化器官的上皮癌细胞系表达的α(1,3/1,4)岩藻糖基转移酶量与唾液酸化路易斯a抗原的表达密切相关。用FTA1-16对结肠癌组织进行免疫组织化学染色显示,与正常细胞相比,癌细胞中该酶的表达增强。最后,使用在大肠杆菌中表达的截短重组肽确定了FTA1-16识别的抗原表位。确定的最小长度是α(1,3/1,4)岩藻糖基转移酶的一个片段,氨基酸位置为132 - 153。