title: Distinct pattern of antibody reactivity with oligomeric or polymeric forms of the capsular polysaccharide of Haemophilus influenzae type b.
abstract: The chain length of oligosaccharides required for antibody binding has been studied by using the capsular polysaccharide from Haemophilus influenzae type b or oligosaccharides derived from it. The concentration of competing antigens required to achieve a 50% inhibition of antibody binding by human polyclonal antisera in an in vitro competition enzyme-linked immunosorbent assay decreased progressively from greater than 10(-3) to 5 x 10(-7) M as the inhibiting saccharide chain length increased from 1 to 262 repeat units. Even small oligosaccharides (one or two repeat units) are potentially capable of competing to a significant level if a high enough concentration of saccharides is used. A similar pattern of reactivity was seen with a monoclonal anti-polyribosyl ribitol phosphate antibody, suggesting that the differences in the avidity of the antibody subpopulations in the polyclonal antisera do not contribute to the binding patterns observed. The binding reaction was specific as evaluated with pneumococcal saccharides. Furthermore, an oligosaccharide-protein conjugate binds antibody better than the free oligosaccharides do. Such a difference in binding was not observed between the polysaccharide and a polysaccharide-protein conjugate. Overall, the data suggest that identical epitopes are expressed by oligomeric and polymeric forms of the antigen and that a particularly more stable conformation in polysaccharides is preferred by antibodies. Covalent coupling of oligomers to protein increases the expression of stable conformation of epitopes. The data further suggest that this kind of antigenic analysis may be important for the design and synthesis of glycoconjugate vaccines.
authors.affiliation: Praxis Biologics, Inc., Rochester, New York 14623.
authors.forename: S
authors.initials: S
authors.lastname: Pillai
authors.forename: S
authors.initials: S
authors.lastname: Ciciriello
authors.forename: M
authors.initials: M
authors.lastname: Koster
authors.forename: R
authors.initials: R
authors.lastname: Eby
mesh_terms: D000907:Antibodies, Bacterial / Q000032:analysis*; D000915:Antibody Affinity; D016667:Bacterial Capsules; D001428:Bacterial Vaccines / Q000276:immunology*; D000939:Epitopes / Q000032:analysis; D006001:Glycoconjugates / Q000276:immunology; D018073:Haemophilus Vaccines; D006193:Haemophilus influenzae / Q000276:immunology*; D006801:Humans; D009844:Oligosaccharides / Q000276:immunology*; D011135:Polysaccharides, Bacterial / Q000276:immunology*; D011487:Protein Conformation
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D000939:Epitopes; D006001:Glycoconjugates; D018073:Haemophilus Vaccines; C044237:Haemophilus influenzae type b polysaccharide vaccine; D009844:Oligosaccharides; D011135:Polysaccharides, Bacterial