A monoclonal antibody defining a binary N-acetyllactosaminyl structure in lactoisooctaosylceramide (IV6Gal beta 1----4GlcNAcnLc6): a useful probe for determining differential glycosylation patterns between normal and transformed human fibroblasts.
Monoclonal antibodies A5 and C6 have been reported previously to recognize developmentally regulated determinants involving N-acetyllactosamine [Fenderson B. A., O'Brien D. A., Millette C. F. and Eddy E. M. (1984) Devl Biol. 103, 117-128]. In the present study, the specificity of these antibodies was determined by solid-phase radioimmunoassay and by thin-layer chromatography immunostaining using purified glycolipid standards. Antibody A5 recognized N-acetyllactosamine (type 2 chain; Gal beta 1----4GlcNAc beta 1----3R), irrespective of branching status. In contrast antibody C6 recognized the binary N-acetyllactosamine structure carried on lactoisooctaosylceramide. Antibody C6 did not react with sialosyl or alpha-galactosyl derivatives of the isooctaosyl structure, including human G10, G8 and bovine G9. Thus, unlike other anti-I antibodies, C6 provides a specific probe for both branching status and absence of terminal chain modification. Monoclonal antibodies A5, C6 and anti-I(Ma) were used to investigate glycosylation changes associated with oncogenic transformation. In contrast to results with lectins, these antibodies preferentially labeled the major glycoproteins of SV40-transformed human embryonic lung fibroblasts, including GP80, GP180, GP200 and GP250. The results suggest that increased expression of unsubstituted polylactosamine core structure at the cell surface follows SV40-transformation.
D000328:Adult; D000606:Amino Sugars / Q000032:analysis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007790:Lactosylceramides / Q000276:immunology
D000606:Amino Sugars; D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D007790:Lactosylceramides; C050870:lactoisooctaosylceramide; C000458:N-acetyllactosamine