title: High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines.
abstract: Long-chain polysialic acid (PSA) is expressed on the vertebrate neural cell adhesion molecule (NCAM) during neuronal plasticity. Its structural similarity to the capsular PSAs of some pathogenic bacteria has hampered the development of polysaccharide vaccines against meningitis. The antibodies formed during immunization require a long epitope for binding, and cross-react with host tissue PSA. The nature of the epitope and possible external effectors involved are still unclear. We have evaluated the interaction of PSA with its antibody mAb735 by surface plasmon resonance. The influences of PSA chain length, pH, temperature, ionic environment, and polyamines were also determined. The antibody binding affinity was found to dramatically increase with PSA chain length. A sub-nanomolar dissociation constant (K(D)=8.5 x 10(-10)M) was obtained for the binding of very long chain native MenB polysaccharides (approximately 200 Neu5Ac-residues). Colominic acid from Escherichia coli K1 (approximately 100 residues) and shorter polymers exhibited progressively weaker affinities. The antibody also bound tightly (K(D) approximately 5 x 10(-9)M) to polysialylated glycopeptides from human embryonal brain. The effects of pH and ionic strength suggested that the interaction is largely electrostatic. Ca2+ and Mn2+ ions promoted the observed surface plasmon resonance response in a concentration dependent fashion. Spermine increased the response in a similar way. Our results suggest that divalent cations and polyamines may play significant role in the regulation of the PSA epitope presentation in vivo.
authors.affiliation: Department of Biochemistry, University of Kuopio, FIN-70211 Kuopio, Finland. jukka.hayrinen@uku.fi
authors.forename: Jukka
authors.initials: J
authors.lastname: Häyrinen
authors.forename: Simon
authors.initials: S
authors.lastname: Haseley
authors.forename: Philippe
authors.initials: P
authors.lastname: Talaga
authors.forename: Martina
authors.initials: M
authors.lastname: Mühlenhoff
authors.forename: Jukka
authors.initials: J
authors.lastname: Finne
authors.forename: Johannes F G
authors.initials: JF
authors.lastname: Vliegenthart
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000915:Antibody Affinity; D016667:Bacterial Capsules; D002118:Calcium / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D009994:Osmolar Concentration; D011073:Polyamines / Q000494:pharmacology; D011135:Polysaccharides, Bacterial / Q000276:immunology*; D012794:Sialic Acids / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D007093:Imidazoles; D011073:Polyamines; D011135:Polysaccharides, Bacterial; D012794:Sialic Acids; C039455:capsular polysaccharide, meningococcal group B; C021319:polysialic acid; C029899:imidazole; D008274:Magnesium; D002118:Calcium