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- W2100047606 abstract "Monoclonal antibodies were prepared against the trisaccharide Galal-3Galpl-4GlcNAc, a sequence which occurs on the surface of Ehrlich ascites tumor cells as well as in thyroglobulin, laminin and a variety of other proteins. This was accomplished by immunizing BALB/c mice with the fraction of Ehrlich cell membrane glyco- proteins obtained by affinity chromatography on a Griffonia simplicifolia I (GS I) column which selectively binds a-D-galactosyl-terminated structures. Detection of Galal-3Galbl-4GlcNAc-specific antibodies was accom- plished by employing glycoproteins containing the trisac- charide sequence; fusion with spleen cells from an immu- nized mouse was accomplished in the presence of polyethylene glycol (PEG1500). An enzyme-linked im- munosorbent assay (ELISA) system was used to identify two clones (2.10G and 6.8E), which recognized the de- sired trisaccharide conjugate. These clones also recog- nized a thyroglobulin fraction isolated by GS I affinity chromatography and murine laminin, both of which pos- sess the Galal-3Galpl-4GlcNAc sequence. Inhibition of antibody-trisaccharide reactivity, examined employing an ELISA assay, revealed that two trisaccharides, Galal-3Galpl-4GlcNAc/Glc, were the best inhibitory haptens; Galpl-4GlcNAc (LacNAc), Galal-3Gal and Galpl-4Glc (lactose) were poor inhibitors. Indirect im- munofluorescence staining of unfixed Ehrlich cells using the monoclonal antibody at 4O C revealed fluorescence over the entire cell surface. Indirect immunogold labeling of semithin and ultrathin sections of aldehyde fixed and Lowicryl K4M-embedded Ehrlich cells resulted in specific labeling of the cell surface and internal structure. Immunoblot analysis revealed that removal of the a- galactosyl residues of laminin by a-galactosidase abol- ished reactivity with the monoclonal antibodies. The availability of this antibody, which belongs to the IgM family of immunoglobulins, now makes possible the de- tection of this sugar sequence on cells and tissue sections, as well as on glycoproteins in solution." @default.
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- W2100047606 date "1993-01-01" @default.
- W2100047606 modified "2023-09-26" @default.
- W2100047606 title "Monoclonal antibodies that recognize V the trisaccharide epitope Galal-3GalS1-4GlcNAc present on ~hrlich tumor cell membrane glycoproteins" @default.
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