Tanimoto Takeshi, Nakatsu Ritsuko, Fuke Isao, Ishikawa Toyokazu, Ishibashi Masahide, Yamanishi Kouichi, Takahashi Michiaki, Tamura Shin-ichi
Laboratory of Prevention of Viral Disease (Research Foundation for Microbial Diseases of Osaka University), Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Vaccine. 2005 Aug 31;23(37):4598-609. doi: 10.1016/j.vaccine.2005.04.042.
The neuraminidase (NA) of the influenza virus, as well as the hemagglutinin, is the most important protective components in the vaccine. However, the NA content of the vaccine remains to be standardized because of the labile nature of this glycoprotein during various chemical treatments and storage. In the present study, the NA content of the split-product (SP) vaccine (virus treated with ether then formalin) was estimated together with that of the virus by an immunochoromatography technique using monoclonal antibodies (mAbs) to viral NA for A/Panama/2007/99 (A/Pa) (H3N2), B/Shangdong/7/97 (B/S) or A/New Caledonia/20/99 (A/NC) (H1N1) viral strains. In the new method, the NA catalytic activity of each fraction from steps of NA purification was measured as an index of NA content. The NA level of A/Pa, B/S or A/NC viral particles was estimated at 6.9+/-0.9, 7.6+/-0.8 or 8.5+/-1.7% of total viral protein (not significant difference between viral strains). The NA level of the corresponding A/Pa, B/S or A/NC vaccines was estimated at 9.6+/-1.5, 12.7+/-0.4 or 12.2+/-1.2% of the total vaccine protein (a significant difference between each strain of virus and its vaccine). These results suggest that the NA content in the N1, N2 or B type NA virus ranges from 5 to 11% of the total viral protein, and that the NA level in each split-product vaccine is 1.4- to 1.6-fold higher than that in the corresponding viral particles. They also suggest that the NA content can be estimated by the immunochoromatography technique using anti-viral NA mAbs.
流感病毒的神经氨酸酶（NA）与血凝素一样，是疫苗中最重要的保护性成分。然而，由于这种糖蛋白在各种化学处理和储存过程中性质不稳定，疫苗中的NA含量仍有待标准化。在本研究中，使用针对A/巴拿马/2007/99（A/Pa）（H3N2）、B/山东/7/97（B/S）或A/新喀里多尼亚/20/99（A/NC）（H1N1）病毒株的病毒NA的单克隆抗体（mAb），通过免疫层析技术对裂解产物（SP）疫苗（经乙醚然后福尔马林处理的病毒）中的NA含量与病毒的NA含量进行了估算。在新方法中，将NA纯化步骤中各组分的NA催化活性作为NA含量的指标进行测定。A/Pa、B/S或A/NC病毒颗粒的NA水平估计分别为总病毒蛋白的6.9±0.9%、7.6±0.8%或8.5±1.7%（病毒株之间无显著差异）。相应的A/Pa、B/S或A/NC疫苗的NA水平估计分别为总疫苗蛋白的9.6±1.5%、12.7±0.4%或12.2±1.2%（每种病毒株与其疫苗之间存在显著差异）。这些结果表明，N1、N2或B型NA病毒中的NA含量占总病毒蛋白的5%至11%，并且每种裂解产物疫苗中的NA水平比相应病毒颗粒中的高1.4至1.6倍。它们还表明，可以使用抗病毒NA单克隆抗体通过免疫层析技术估算NA含量。
