Hanisch F G, Heimbüchel G, Baldus S E, Uhlenbruck G, Schmits R, Pfreundschuh M, Schwonzen M, Vierbuchen M, Bara J, Peter-Katalinic J
Institute of Immunobiology, University of Cologne, Federal Republic of Germany.
Cancer Res. 1993 Sep 15;53(18):4367-75.
A mouse IgM monoclonal antibody FW6 was established after immunization of mice with mucins from human amniotic fluid and was characterized with regard to its binding epitope. According to a series of biochemical criteria the epitope is located on O-linked neutral carbohydrates of M(r) 700,000 and M(r) 570,000 mucins in human amniotic fluid. The epitope is presumed to contain alpha 3/4-linked fucose and terminal beta 3/4-linked galactose that are labile to the fucosidase I from almond emulsion or to the galactosidase from bovine testes, respectively. Immunoreactive fractions of glycan alditols from human amniotic fluid mucins were partially characterized by fast atom bombardment-mass spectrometry and methylation analysis to be composed of monofucosylated polylactosamine-type deca- or nonasaccharides. According to antibody competition studies, inhibition assays with defined carbohydrates and binding assays on neoglycolipids monoclonal antibody FW6 are presumed to recognize a novel epitope that is distinct from known carbohydrate markers of the Lex/Ley family associated with colonic carcinomas. The selective reactivity of this monoclonal antibody to the majority of human colonic carcinomas and its nonreactivity to normal colonic mucosa may render this antibody as a valuable tool in cancer diagnosis or cancer treatment.
用来自人羊水的粘蛋白免疫小鼠后建立了小鼠IgM单克隆抗体FW6，并对其结合表位进行了表征。根据一系列生化标准，该表位位于人羊水分子量为700,000和570,000的粘蛋白的O-连接中性碳水化合物上。推测该表位含有分别对杏仁乳液中的岩藻糖苷酶I或牛睾丸中的半乳糖苷酶不稳定的α3/4连接的岩藻糖和末端β3/4连接的半乳糖。通过快速原子轰击质谱和甲基化分析对人羊水粘蛋白的聚糖糖醇免疫反应性组分进行了部分表征，其由单岩藻糖基化的聚乳糖胺型十糖或九糖组成。根据抗体竞争研究、用特定碳水化合物进行的抑制试验以及对新糖脂的结合试验，推测单克隆抗体FW6识别一种与已知的与结肠癌相关的Lex/Ley家族碳水化合物标记不同的新表位。这种单克隆抗体对大多数人结肠癌的选择性反应及其对正常结肠粘膜的无反应性可能使其成为癌症诊断或癌症治疗中的一种有价值的工具。
