title: Characterization of size, structure and purity of serogroup X Neisseria meningitidis polysaccharide, and development of an assay for quantification of human antibodies.
abstract: Serogroup X Neisseria meningitidis (MenX) has recently emerged as a cause of localized disease outbreaks in sub-Saharan Africa. In order to prepare for vaccine development, MenX polysaccharide (MenX PS) was purified by standard methods and analyzed for identity and structure by NMR spectroscopy. This study presents the first full assignment of the structure of the MenX PS using (13)C, (1)H and (31)P NMR spectroscopy and total correlation spectroscopy (TOCSY) and (1)H-(13)C heteronuclear single quantum coherence (HSQC). Molecular size distribution analysis using HPLC-SEC with multi-angle laser light scattering (MALLS) found the single peak of MenX PS to have a weight-average molar mass of 247,000g/mol, slightly higher than a reference preparation of purified serogroup C meningococcal polysaccharide. MenX PS tended to be more thermostable than serogroup A PS. A method for the quantification of MenX PS was developed by use of high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). A novel and specific ELISA assay for quantification of human anti-MenX PS IgG based on covalent linkage of the MenX PS to functionally modified microtitre plates was developed and found valid for the assessment of the specific antibody concentrations produced in response to MenX vaccination or natural infection. The current work thus provides the necessary background for the development of a MenX PS-based vaccine to prevent meningococcal infection caused by bacteria bearing this capsule.
authors: Oxford Vaccine Group, Department of Paediatrics, University of Oxford, and the NIHR Oxford Biomedical Research Centre, Oxford OX3 7TU, UK.
authors: Ouli
authors: O
authors: Xie
authors: Barbara
authors: B
authors: Bolgiano
authors: Fang
authors: F
authors: Gao
authors: Kay
authors: K
authors: Lockyer
authors: Carolyn
authors: C
authors: Swann
authors: Christopher
authors: C
authors: Jones
authors: Isabelle
authors: I
authors: Delrieu
authors: Berthe-Marie
authors: BM
authors: Njanpop-Lafourcade
authors: Tsidi Agbeko
authors: TA
authors: Tamekloe
authors: Andrew J
authors: AJ
authors: Pollard
authors: Gunnstein
authors: G
authors: Norheim
mesh_terms: D000293:Adolescent; D000328:Adult; D017954:Africa South of the Sahara; D000907:Antibodies, Bacterial / Q000276:immunology*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D058977:Molecular Sequence Annotation; D015394:Molecular Structure; D009345:Neisseria meningitidis / Q000737:chemistry* / Q000145:classification / Q000276:immunology; D011135:Polysaccharides, Bacterial / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D012703:Serotyping; D055815:Young Adult
chemical_list: D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D011135:Polysaccharides, Bacterial