Sialic acid polysaccharide antigens of Neisseria meningitidis and Escherichia coli: esterification between adjacent residues

Abstract

Colominic acid and meningococcal Group B polysaccharide, both (2 leads to 8)-alpha-linked homopolymers of sialic acid, are made water-insoluble either by reaction with a carbodiimide in aqueous solution at pH 4.75 or by treatment with 48% aqueous hydrofluoric acid at 0 degrees for 48 h. I.r. spectra of the products show a major band near 1750 cm-1, consistent with ester formation; this band virtually disappears after mild, alkali treatment. Esterification also occurs by incubating the native polysaccharides below pH 6.0, and their i.r. spectra show that the degree of esterification increases as the pH is lowered. The relatively low molecular weight of these partially esterified, water-soluble polymers is consistent with intra- rather than inter-molecular ester formation. Counter-current immunoelectrophoresis shows that esterification of approximately 9% is sufficient to abolish immunoprecipitation. 13C-N.m.r. spectroscopy of fully esterified colominic acid provides strong evidence for cross-linking between the carboxyl group of one residue and HO-9 of an adjoining residue. Meningococcal serogroup C polysaccharide, a (2 leads to 9)-alpha-linked homopolymer of sialic acid containing O-acetyl groups at C-7 and/or C-8, does not undergo intramolecular esterification after carbodiimide treatment. However, upon O-deacetylation of the polysaccharide, esterification occurs.