Elsevier

Analytical Biochemistry

Volume 442, Issue 2, 15 November 2013, Pages 259-261
Analytical Biochemistry

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Meningococcal X polysaccharide quantification by high-performance anion-exchange chromatography using synthetic N-acetylglucosamine-4-phosphate as standard

https://doi.org/10.1016/j.ab.2013.08.001Get rights and content

Abstract

A method for meningococcal X (MenX) polysaccharide quantification by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC–PAD) is described. The polysaccharide is hydrolyzed by strong acidic treatment, and the peak of glucosamine-4-phosphate (4P-GlcN) is detected and measured after chromatography. In the selected conditions of hydrolysis, 4P-GlcN is the prevalent species formed, with GlcN detected for less than 5% in moles. As standard for the analysis, the monomeric unit of MenX polysaccharide, N-acetylglucosamine-4-phosphate (4P-GlcNAc), was used. This method for MenX quantification is highly selective and sensitive, and it constitutes an important analytical tool for the development of a conjugate vaccine against MenX.

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Acknowledgments

We thank Gerd Pluschke of the Swiss Tropical Institute (Basel, Switzerland), who kindly provided the Neisseria meningitidis X5967 strain (ST 750) used for MenX polysaccharide production by bacterial growth.

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  • Quantitation of novel pentavalent meningococcal polysaccharide conjugate vaccine (Men A-TT, Men C-CRM, Men Y-CRM, Men W-CRM, Men X-TT) using sandwich ELISA

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    Available meningococcal vaccines are unable to provide protection against serogroup X [17] and currently no vaccine is available in the market against Men X [18]. However, recent development of a glycoconjugate vaccine against Men X has been reported [11,16]. World Health Organization’s increasing interest towards inclusion of serogroup X as one of the causative agents for epidemic meningitis [19], has raised a challenge for vaccine industry to develop efficient vaccine against Men X.

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