A monoclonal antibody reactive with terminal lactotriaosyl residue-containing oligosaccharides and its application to characterizing cell surface expression of the glyco-epitopes in COS-1 cells.
A new monoclonal antibody (MoAb), designated JF12, reactive strongly with Lc3 (GlcNAc beta 1-->3Gal beta 1-->4Glc beta 1-->Cer) but only slightly with nLc5 (GlcNAc beta 1-->3Gal beta 1-->4GlcNAc beta 1-->3Gal beta 1-->4Glc beta 1-->Cer) has been prepared after immunization of Balb/c mice with Lc3. By the flowcytometrical analyses of COS-1 cells, which was strongly stained with JF12, the reactivity completely disappeared in the confluent condition upon harvesting by the protease treatments of the cell surface. In the sparse condition, however, the cells still retained JF12 reactivity in spite of the protease treatments. This strongly suggests that the expression of terminal GlcNAc-containing glycoconjugates on the surface of COS-1 cells may be dramatically modulated by protease-sensitive membranous components being dependent upon the cell density.
Nakamura
M
M
Division of Hemopoiesis, Jichi Medical School, Tochigi, Japan.
Tsunoda
A
A
Sakoe
K
K
Saito
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D010450:Endopeptidases; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006020:Glycopeptides / Q000032:analysis / Q000096:biosynthesis*; D007790:Lactosylceramides / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D014162:Transfection
D000911:Antibodies, Monoclonal; D006020:Glycopeptides; D007790:Lactosylceramides; D009844:Oligosaccharides; C025575:lactotriaosylceramide; C085206:N-acetylglucosaminyl-beta-1-3-galactosyl-1-4-(N-acetylglucosaminyl)-1-3-galactosyl-1-4-glucosyl-1-ceramide; D010450:Endopeptidases