Meningococcal X polysaccharide quantification by high-performance anion-exchange chromatography using synthetic N-acetylglucosamine-4-phosphate as standard.
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.
Novartis Vaccines Institute for Global Health, Via Fiorentina 1, I-53100 Siena, Italy.
F
F
Micoli
R
R
Adamo
D
D
Proietti
M
M
Gavini
M R
MR
Romano
C A
CA
MacLennan
P
P
Costantino
F
F
Berti
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards*; D002852:Chromatography, Ion Exchange / Q000379:methods* / Q000592:standards*; D004563:Electrochemistry; D005958:Glucosephosphates / Q000032:analysis*; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D012015:Reference Standards
D005958:Glucosephosphates; D011135:Polysaccharides, Bacterial; D000117:Acetylglucosamine
Carbohydrate-based vaccine; Meningococcal serogroup X polysaccharide; Quantitative estimation