An effective method for the synthesis of neoglycoproteins and neogangliosideproteins by use of reductively aminated sulfhydryl-containing carbohydrate conjugates.
A highly effective method for the synthesis of neoglycoconjugates has been developed. The technique is based on two steps. First, a sulfhydryl group is introduced into the carbohydrate by reductive amination with cysteamine, and secondly the sulfhydryl group carrying carbohydrate is coupled to a protein or carrier via a disulfide bridge. The method was used to couple sialic acid-containing carbohydrates such as N-acetylneuraminyllactose and the carbohydrate moieties of gangliosides (GM1, GM3 and GD1b) to bovine serum albumin. Biological functionality of the generated GM1- and GD1b-neoglycoprotein was demonstrated by a choleratoxin-binding assay and an immunoassay, respectively. The applicability of neoglycoconjugates for the quantification of gangliosides in solution by a competitive assay is shown.
Schneller
M
M
Abteilung für Klinische Chemie, Universität München.
Geiger
R E
RE
D000586:Amination; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001667:Binding, Competitive; D002772:Cholera Toxin / Q000737:chemistry; D003543:Cysteamine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005732:Gangliosides / Q000032:analysis / Q000138:chemical synthesis* / Q000276:immunology; D006001:Glycoconjugates / Q000737:chemistry*; D006023:Glycoproteins / Q000032:analysis / Q000138:chemical synthesis*; D006801:Humans; D009844:Oligosaccharides / Q000737:chemistry; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012996:Solutions; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D004220:Disulfides; D005732:Gangliosides; D006001:Glycoconjugates; D006023:Glycoproteins; D009844:Oligosaccharides; D012996:Solutions; D013438:Sulfhydryl Compounds; D012710:Serum Albumin, Bovine; D003543:Cysteamine; D002772:Cholera Toxin