Meningococcal X polysaccharide quantification by high-performance anion-exchange chromatography using synthetic N-acetylglucosamine-4-phosphate as standard |
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| Authors: | F Micoli R Adamo D Proietti M Gavini MR Romano CA MacLennan P Costantino F Berti |
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| Institution: | 1. Novartis Vaccines Institute for Global Health, Via Fiorentina 1, I-53100 Siena, Italy;2. Novartis Vaccines and Diagnostics Research, Via Fiorentina 1, I-53100 Siena, Italy;3. MRC Centre for Immune Regulation, School of Immunity and Infection, College of Medicine and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK |
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| 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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| Keywords: | Meningococcal serogroup X polysaccharide Quantitative estimation Carbohydrate-based vaccine |
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