Ravenscroft N, Averani G, Bartoloni A, Berti S, Bigio M, Carinci V, Costantino P, D'Ascenzi S, Giannozzi A, Norelli F, Pennatini C, Proietti D, Ceccarini C, Cescutti P
Chiron Vaccines SpA, Siena, Italy.
Vaccine. 1999 Jul 16;17(22):2802-16. doi: 10.1016/s0264-410x(99)00092-4.
We recently described the use of ion exchange chromatography for analysis and the industrial scale preparation of pools of oligosaccharides of intermediate chain length from polysaccharides of Haemophilus influenzae type b (Hib) and Neisseria meningitidis groups A and C. These negatively charged "sized" oligosaccharides are activated and conjugated to the carrier protein (CRM197) to prepare the corresponding glycoconjugate vaccines. Characterization and accurate determination of the degree of polymerization (DP) of the pool of oligosaccharides is essential for the consistent production of these conjugate vaccines. This paper describes the colorimetric assays used for determination of the average DP of the Hib and meningococcal oligosaccharides, and the qualification of these assays achieved by size characterization of the respective oligosaccharides by use of physicochemical methods, including liquid chromatography, mass spectrometry (ionspray) and NMR spectroscopy.
我们最近描述了使用离子交换色谱法分析和工业规模制备来自b型流感嗜血杆菌（Hib）以及A群和C群脑膜炎奈瑟菌多糖的中等链长寡糖库。这些带负电荷的“分级”寡糖被活化并与载体蛋白（CRM197）偶联，以制备相应的糖共轭疫苗。寡糖库聚合度（DP）的表征和准确测定对于这些共轭疫苗的稳定生产至关重要。本文描述了用于测定Hib和脑膜炎球菌寡糖平均DP的比色测定法，以及通过使用包括液相色谱、质谱（离子喷雾）和核磁共振光谱在内的物理化学方法对相应寡糖进行尺寸表征而实现的这些测定法的鉴定。
