Size determination of bacterial capsular oligosaccharides used to prepare conjugate vaccines against Neisseria meningitidis groups Y and W135.
The glycoconjugate vaccines against Neisseria meningitidis groups Y and W135 consist of pools of selected oligosaccharides conjugated to the protein carrier (CRM197). Consistent production of these vaccines requires control and thus determination of the average degree of polymerisation of the oligosaccharides used for conjugation. Acid hydrolysis generates group Y and W135 oligosaccharides with N-acetylneuraminic acid at the reducing end. A method, involving NaBH4 reduction and quantification of this terminal N-acetylneuraminic acid by use of high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) following acid hydrolysis (2M TFA), was developed. The average degree of polymerisation is calculated from the ratio of reduced N-acetylneuraminic acid to total N-acetylneuraminic acid. The assay was qualified by application to group C, Y and W135 oligosaccharide standards characterised by liquid chromatography, mass and NMR spectroscopy.
Bardotti
Angela
A
Chiron Vaccines, Via Fiorentina 1, I-53100 Siena, Italy. angela_bardotti@chiron.it
Averani
Giovanni
G
Berti
Francesco
F
Berti
Stefania
S
Galli
Chiara
C
Giannini
Sara
S
Fabbri
Barbara
B
Proietti
Daniela
D
Ravenscroft
Neil
N
Ricci
Stefano
S
D002240:Carbohydrate Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D006868:Hydrolysis; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D022401:Meningococcal Vaccines / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009345:Neisseria meningitidis / Q000145:classification / Q000276:immunology*; D009844:Oligosaccharides / Q000737:chemistry* / Q000276:immunology*; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization; D018074:Vaccines, Conjugate / Q000737:chemistry / Q000276:immunology
D007202:Indicators and Reagents; D022401:Meningococcal Vaccines; D009844:Oligosaccharides; D011134:Polysaccharides; D018074:Vaccines, Conjugate; D019158:N-Acetylneuraminic Acid