McIlhinney R A, Patel S, Gore M E
Biochem J. 1985 Apr 1;227(1):155-62. doi: 10.1042/bj2270155.
The molecules of the human milk fat globule membrane (MFGM) which bind four murine monoclonal antibodies (LICR LON M3, M8, M18 and M24) raised against the human MFGM have been identified. By using 'Western' blotting [Burnette (1981) Anal. Biochem. 112, 195-203] it was shown that each antibody reacted with a different set of proteins. M3 and M24 were similar in their pattern of reaction with the membrane proteins, but were quite distinct from M8 and M18, which also differed from each other. Glycopeptides prepared from the MFGM by exhaustive Pronase digestion were able to inhibit partially the binding of M3 and M24, and prevent totally the binding of M8 and M18, to the MFGM in an enzyme-linked immunoabsorbent assay. Oligosaccharides obtained by the deproteination of human milk also completely inhibited the binding of M3, M18 and M24 to the MFGM. However, the binding of M8 was not inhibited by these saccharides, and therefore M8 may not be recognizing a simple carbohydrate determinant. By using an enzyme-linked assay, M8 and M18 were shown not to bind to MFGM glycolipid, whereas M3 and M24 did, and this was confirmed by overlaying thin layer chromatograms of MFGM lipids with these antibodies. Both M3 and M24 showed a similar complex pattern of reaction, binding to more than one glycolipid moiety. By these means all four antibodies have been shown to react with antigens which involve carbohydrate side chains carried on different proteins, and two were also shown to react with such determinants on glycolipids.
已鉴定出与人乳脂肪球膜（MFGM）结合的4种抗人MFGM的鼠单克隆抗体（LICR LON M3、M8、M18和M24）的分子。通过“Western”印迹法[Burnette（1981年）《分析生物化学》112卷，第195 - 203页]表明，每种抗体与不同的蛋白质组发生反应。M3和M24与膜蛋白的反应模式相似，但与M8和M18截然不同，而M8和M18彼此之间也存在差异。在酶联免疫吸附测定中，通过用链霉蛋白酶彻底消化从MFGM制备的糖肽能够部分抑制M3和M24与MFGM的结合，并完全阻止M8和M18与MFGM的结合。通过人乳脱蛋白获得的寡糖也完全抑制了M3、M18和M24与MFGM的结合。然而，这些糖类并未抑制M8的结合，因此M8可能识别的不是简单的碳水化合物决定簇。通过酶联测定表明，M8和M18不与MFGM糖脂结合，而M3和M24则与之结合，用这些抗体覆盖MFGM脂质的薄层色谱图证实了这一点。M3和M24都表现出相似的复杂反应模式，与不止一种糖脂部分结合。通过这些方法已表明，所有这4种抗体都与涉及不同蛋白质上携带的碳水化合物侧链的抗原发生反应，并且还表明其中两种抗体也与糖脂上的此类决定簇发生反应。
