Ricci S, Bardotti A, D'Ascenzi S, Ravenscroft N
Chiron Vaccines SpA, Via Fiorentina 1, I-53100, Siena, Italy.
Vaccine. 2001 Feb 28;19(15-16):1989-97. doi: 10.1016/s0264-410x(00)00427-8.
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.
已开发出一种定量测定A群脑膜炎奈瑟菌（MenA）荚膜多糖（CPS）的新方法。该方法基于对CPS进行三氟乙酸（TFA）水解（在80℃下2M水解3小时），然后进行色谱分离，并使用与灵敏脉冲安培检测器ED40联用的IonPac AS11柱，从所得峰面积中对释放的6-磷酸甘露糖胺进行定量。该方法的高选择性避免了基于磷比色测定的经典方法所存在的干扰问题。只要能找到合适的水解条件，这种色谱方法可能适用于其他含磷酸化糖的细菌抗原的定量，如H、L、X和Z群脑膜炎球菌，以及6、10A和19型肺炎球菌血清型。
