Quantitative determination of saccharide in Haemophilus influenzae type b glycoconjugate vaccines, alone and in combination with DPT, by use of high-performance anion-exchange chromatography with pulsed amperometric detection

被引：27

作者：

Bardotti, A ^{[1
]}

Ravenscroft, N ^{[1
]}

Ricci, S ^{[1
]}

D'Ascenzi, S ^{[1
]}

Guarnieri, V ^{[1
]}

Averani, G ^{[1
]}

Constantino, P ^{[1
]}

机构：

[1] Chiron Vaccines SpA, I-53100 Siena, Italy

来源：

VACCINE | 2000年 / 18卷 / 19期

关键词：

conjugate vaccines; Haemophilus influenzae type b; saccharide analysis; HPAEC-PAD;

D O I：

10.1016/S0264-410X(99)00535-6

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

The stability and integrity of glycoconjugate vaccines requires determination of the total saccharide and quantification of the unbound or free saccharide present. The traditional assay for Hib conjugates, based on colorimetric determination of ribose, has been much improved by the use of base hydrolysis and analysis of the Hib subunit generated using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The production of this subunit was confirmed by NMR analysis. However, quantification of free Hib saccharide using this method was not possible in the combination vaccines evaluated due to interferences emanating from DPT. Thus a method based on TFA hydrolysis followed by the chromatographic separation and quantification of ribitol on a CarboPac MAI column was developed. The method is selective, and with the use of ED40 electrode, requires only nanomole amounts for the chromatographic step, thereby ensuring that free saccharide can be monitored accurately in the formulated Hib-CRM vaccine alone and when in combination with other vaccines. (C) 2000 Elsevier Science Ltd. All rights reserved.

引用

页码：1982 / 1993

页数：12

共 19 条

[1]

ADAMS WG, 1992, 32 ICAAC AN CA, P273

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