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.

Original publication

DOI

10.1016/j.vaccine.2012.07.032

Type

Journal article

Journal

Vaccine

Publication Date

31/08/2012

Volume

30

Pages

5812 - 5823

Keywords

Adolescent, Adult, Africa South of the Sahara, Antibodies, Bacterial, Chromatography, Gel, Chromatography, High Pressure Liquid, Enzyme-Linked Immunosorbent Assay, Humans, Immunoglobulin G, Magnetic Resonance Spectroscopy, Molecular Sequence Annotation, Molecular Structure, Neisseria meningitidis, Polysaccharides, Bacterial, Serotyping, Young Adult