Development of a new method for the quantitative analysis of the extracellular polysaccharide of Neisseria meningitidis serogroup A by use of high-performance anion-exchange chromatography with pulsed-amperometric detection.
A new method for the quantitative determination of Neisseria meningitidis group A (MenA) capsular polysaccharide (CPS) has been developed. The method is based on trifluoracetic acid (TFA) hydrolysis of the CPS (2 M at 80 degrees C for 3 h), followed by chromatographic separation and quantification of the liberated mannosamine-6-phosphate from the area of the peak obtained using an IonPac AS11 column coupled to the sensitive pulsed amperometric detector ED40. The highly selective nature of this method circumvents the interference problems associated with the classical method based on a colorimetric assay for phosphorus. Provided that suitable hydrolysis conditions can be found, this chromatographic approach might be applicable to the quantification of other bacterial antigens containing phosphorylated sugars such as meningococcal groups H, L, X and Z, and pneumococcal serotypes 6, 10A and 19.
Chiron Vaccines SpA, Via Fiorentina 1, I-53100, Siena, Italy. stefano_ricci@chiron.it
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Ricci
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Bardotti
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D'Ascenzi
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Ravenscroft
D002852:Chromatography, Ion Exchange / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D022401:Meningococcal Vaccines / Q000737:chemistry; D015394:Molecular Structure; D009345:Neisseria meningitidis / Q000737:chemistry / Q000145:classification / Q000276:immunology*; D011135:Polysaccharides, Bacterial / Q000032:analysis* / Q000737:chemistry; D012703:Serotyping; D014269:Trifluoroacetic Acid; D018074:Vaccines, Conjugate / Q000737:chemistry
D022401:Meningococcal Vaccines; D011135:Polysaccharides, Bacterial; D018074:Vaccines, Conjugate; D014269:Trifluoroacetic Acid