HPLC-based quantification of haemagglutinin in the production of egg- and MDCK cell-derived influenza virus seasonal and pandemic vaccines

被引：51

作者：

Kapteyn, J. C. ^{[1
]}

Porre, A. M. ^{[1
]}

de Rond, E. J. R. ^{[1
]}

Hessels, W. B. ^{[1
]}

Tijms, M. A. ^{[1
]}

Kessen, H. ^{[1
]}

Slotboom, A. M. E. ^{[1
]}

Oerlemans, M. A. ^{[1
]}

Smit, D. ^{[1
]}

van der Linden, J. ^{[1
]}

Schoen, P. ^{[1
]}

Thus, J. L. G. ^{[1
]}

机构：

[1] Solvay Biol, Biopharmaceut R&D, NL-1381 CP Weesp, Netherlands

来源：

VACCINE | 2009年 / 27卷 / 09期

关键词：

Haemagglutinin; SRID; H5N1; Influenza vaccine; SINGLE-RADIAL-IMMUNODIFFUSION; PERFORMANCE LIQUID-CHROMATOGRAPHY; ASSAY;

D O I：

10.1016/j.vaccine.2008.11.113

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 haemagglutinin (HA) content is an important specification of influenza vaccines. Recently, a reversed-phase high performance liquid chromatography (RP-HPLC) method for quantification of HA in PER.C6 (R) cell culture-based whole virus vaccines has been reported, having a high sensitivity, precision, broad range, and high sample throughput [Kapteyn JC, Drissi Saidi M, Dijkstra R, Kars C, Tjon Cm-K, Weverling GJ et al. Haemagglutinin quantification and identification of influenza A&B strains propagated in PER.C6 (R) cells: a novel RP-HPLC method. Vaccine 2006:24:3137-44]. This RP-HPLC assay is based on measuring the peak area of HA1, the hydrophilic subunit of HA, which turned out to be proportional to the amount of HA analyzed. Here, we present data demonstrating that this RP-HPLC method is also highly suitable for HA quantification of active and BPL- or formaldehyde-inactivated egg-based and MDCK cell-based whole virus samples, including egg allantoic harvest, and in final (monovalent) subunit vaccines, including those for pandemic H5N1 strains and for virosomal vaccines. In addition, the RP-HPLC assay was demonstrated to be a very powerful tool in the early stages of seasonal influenza vaccine production, when homologous serial radial immunodiffusion (SRID) reagents are not yet available, enabling fast and reliable viral growth studies in eggs in order to select the best growing virus strains or reassortants for the production of the seasonal trivalent influenza vaccine. Because of its high sensitivity, the RP-HPLC assay has shown its enormous value in supporting small scale MDCK-based (H5N1) influenza virus production models. Finally, the observed differences between HAI molecules from various HA subtypes in UV absorbance, FLD response, and in the actual retention times in RP-HPLC are discussed in relation to the primary structure of the HAI molecules studied. (C) 2008 Elsevier Ltd. All rights reserved.

引用

页码：1468 / 1477

页数：10

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