Marcus D M, Gilbert S, Sekine M, Suzuki A
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.
Arch Biochem Biophys. 1988 May 1;262(2):620-5. doi: 10.1016/0003-9861(88)90414-6.
The 3-fucosyllactosamine (3-FL) antigenic determinant, Gal(beta 1-4)[Fuc(alpha 1-3)]-GlcNAc-R, is very immunogenic in BALB/c mice. A panel of monoclonal antibodies was raised against a novel glycosphingolipid that contains a terminal 3-FL structure (structure 1): (formula: see text) Surprisingly, most of these antibodies reacted with the internal galactosylgloboside structure rather than with the terminal 3-FL epitope. The specificities of two of these antibodies, 5A3 and 8A7, were analyzed by enzyme-linked immunosorbent assay and immunostaining procedures, and compared with MC631 (anti-SSEA-3), the only other monoclonal antibody known to bind to galactosylgloboside. 5A3 and 8A7 bound equally well to structure 1 and to galactosylgloboside. These antibodies also bound to structure 1 from which the terminal galactose, or the galactose and fucose, had been removed, but not to globoside or sialylated galactosylgloboside. In contrast, MC631 did not bind to structure 1 but, as described previously, did bind to globoside and sialylated galactosylgloboside. Galactosylgloboside is a developmentally regulated antigen of mouse embryos, human teratocarcinomas, and mouse embryonic brain. Antibodies 5A3 and 8A7 will complement MC631 in the analysis of the distribution and regulation of galactosylgloboside.
3-岩藻糖基乳糖胺（3-FL）抗原决定簇，即Gal(β1-4)[Fuc(α1-3)]-GlcNAc-R，在BALB/c小鼠中具有很强的免疫原性。针对一种含有末端3-FL结构的新型糖鞘脂（结构1）制备了一组单克隆抗体：（分子式：见正文）令人惊讶的是，这些抗体中的大多数与内部半乳糖基球蛋白结构反应，而不是与末端3-FL表位反应。通过酶联免疫吸附测定和免疫染色程序分析了其中两种抗体5A3和8A7的特异性，并与MC631（抗SSEA-3）进行了比较，MC631是已知的唯一一种与半乳糖基球蛋白结合的单克隆抗体。5A3和8A7与结构1和半乳糖基球蛋白的结合效果相同。这些抗体也与去除了末端半乳糖或半乳糖和岩藻糖的结构1结合，但不与球蛋白或唾液酸化半乳糖基球蛋白结合。相比之下，MC631不与结构1结合，但如先前所述，它确实与球蛋白和唾液酸化半乳糖基球蛋白结合。半乳糖基球蛋白是小鼠胚胎、人类畸胎瘤和小鼠胚胎脑的一种受发育调控的抗原。抗体5A3和8A7将在半乳糖基球蛋白的分布和调控分析中补充MC631。
