甲型H1N1流感疫苗神经氨酸酶含量测定参考品的建立

发布时间：2019-01-26 20:47

【摘要】：目的建立甲型H1N1流感疫苗神经氨酸酶含量测定参考品。方法对甲型H1N1流感疫苗原液进行还原电泳后,采用免疫印迹,糖蛋白染色,方法初步确定甲流H1N1疫苗原液中神经氨酸酶SDS-PAGE条带位置,切取条带后通过edman N端测序法进行确认。采用Lowry法进行总蛋白定量,SDS-PAGE密度扫描的方法确定神经氨酸酶比例,计算出疫苗原液中神经氨酸酶含量。结果确定甲流疫苗原液中神经氨酸酶在SDS-PAGE中相对分子质量约71 000,对多批次疫苗原液进行测定后,选取神经氨酸酶含量较高的批次进行测定,总蛋白质量浓度为1046.00μg/m L,神经氨酸酶质量分数11.78%。二者相乘得出该批次疫苗原液神经氨酸酶含量为123.22μg/m L。结论研究成功建立了甲型H1N1流感疫苗神经氨酸酶含量测定参考品,其他毒株生产的流感疫苗也可以参考该方法建立NA含量测定参考品。
[Abstract]:Objective to establish a reference for the determination of neuraminidase in influenza A H1N1 vaccine. Methods after electrophoretic reduction of influenza A H1N1 vaccine, the neuraminidase SDS-PAGE bands in the H1N1 vaccine solution were preliminarily determined by immunoblotting and glycoprotein staining. The bands were sequenced and confirmed by edman N-terminal sequencing. The Lowry method was used to quantify the total protein and the SDS-PAGE density scanning method was used to determine the proportion of neuraminidase, and the content of neuraminidase in the vaccine solution was calculated. The results showed that the relative molecular weight of neuraminidase in SDS-PAGE was about 71,000. After the determination of the original solution of multiple batches of vaccine, the batches with higher content of neuraminidase were selected to determine the content of neuraminidase. The total protein concentration was 1046.00 渭 g / mL and the neuraminidase mass fraction was 11.78 渭 g / mL. The results showed that the neuraminidase content of the original solution of the vaccine was 123.22 渭 g / mL. Conclusion the reference material for the determination of neuraminidase in influenza A H1N1 vaccine was successfully established, and the reference material for the determination of NA content in influenza vaccine produced by other strains was also established.
【作者单位】：中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室;
【基金】：科技部国际合作专项(2013DFA31680)
【分类号】：R392

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