N. Ravenscroft et al., Size determination of bacterial capsular oligosaccharides used to prepare conjugate vaccines, VACCINE, 17(22), 1999, pp. 2802-2816
We recently described the use of ion exchange chromatography for analysis a
nd the industrial scale preparation of pools of oligsaccharides of intermed
iate chain length from polysaccharides of Haemophilus influenzae of 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. Characteriz
ation and accurate determination of the degree of polymerization (DP) of th
e pool of oligosaccharides is essential for the consistent production of th
ese conjugate vaccines. This paper describes the colorimetric assays used f
or determination of the average DP of the Hib and meningococcal oligosaccha
rides, and the qualification of these assays achieved by size characterizat
ion of the respective oligosaccharides by use of physicochemical methods, i
ncluding liquid chromatography. mass spectrometry (ionspray) and NMR spectr
oscopy. (C) 1999 Elsevier Science Ltd. All rights reserved.