Quantitation of haemagglutinin in H5N1 influenza viruses reveals low haemagglutinin content of vaccine virus NIBRG-14 (H5N1)
Quantitation of haemagglutinin in H5N1 influenza viruses reveals low haemagglutinin content of vaccine virus NIBRG-14 (H5N1)
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DOI：
10.1016/j.vaccine.2008.09.050
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发表时间：
2008-12-02
期刊：
影响因子：
5.5
通讯作者：
Engelhardt, Othmar G.
中科院分区：
文献类型：
--
作者：
Harvey, Ruth;Wheeler, Jun X.;Engelhardt, Othmar G.
对潜在候选流感疫苗病毒的评估包括许多因素。病毒的生长特性和抗原的产量，特别是血凝素(HA)是至关重要的。最近开发的H5N1候选疫苗病毒NIBRG-14(其HA和NA基因来自A/波多黎各/8/34背景的分支1病毒A/越南/1194/2004)被认为产生较低的抗原量。在研究H5N1疫苗病毒的抗原产量时，我们发现由于蛋白质在SDS-PAGE凝胶上的迁移特性，准确定量某些H5N1病毒的HA含量是困难的。HA1和HA2的手分别与核蛋白(NP)和基质蛋白(M1)共同迁移，阻碍了准确的分析。我们已经开发出一种准确的方法，通过在标准方案中引入脱糖步骤来定量这些H5N1病毒的HA。使用这种方法，我们重复地证明了NIBRG-14的低产量至少部分是由于病毒制剂中HA的含量低于正常水平。亲本野生型A/越南/1194/2004病毒也是如此。(C)2008爱思唯尔有限公司。保留所有权利。
The assessment of potential candidate influenza vaccine viruses includes a number of factors. Growth properties of the Virus and yield of antigen, specifically the haemagglutinin (HA), are of key importance. The recently developed H5N1 candidate vaccine Virus NIBRG-14 (with HA and NA genes derived from the clade 1 virus A/Viet Nam/1194/2004 in an A/Puerto Rico/8/34 background) has been Suggested to yield low amounts of antigen. While investigating the antigen yield of H5N1 vaccine Viruses, we found that accurate quantitation of the HA content of some H5N1 Viruses was difficult due to the migration characteristics of the proteins on SDS-PAGE gels. The HA1 and HA2 hands co-migrated with nucleoprotein (NP) and matrix Protein (M1) respectively, preventing accurate analysis. We have developed an accurate way of quantitating HA from these H5N1 viruses by introducing a deglycosylation step to the standard protocol. Using this method, we showed reproducibly that the low yield of NIBRG-14 is, at least in part, due to a lower than usual content of HA in Virus preparations. This was also found to be the case for the parent wild type A/Viet Nam/1194/2004 virus. (C) 2008 Elsevier Ltd. All rights reserved.