Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane.
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.
McIlhinney
R A
RA
Patel
S
S
Gore
M E
ME
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002241:Carbohydrates / Q000302:isolation & purification; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D000939:Epitopes / Q000276:immunology*; D006017:Glycolipids / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007122:Immunoelectrophoresis; D008894:Milk Proteins / Q000276:immunology; D008895:Milk, Human / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D006017:Glycolipids; D006023:Glycoproteins; D008894:Milk Proteins