Label-free mass spectrometry-based quantification of hemagglutinin and neuraminidase in influenza virus preparations and vaccines.
Label-free mass spectrometry-based quantification of hemagglutinin and neuraminidase in influenza virus preparations and vaccines.
复制标题
在流感病毒制剂和疫苗中基于无标记的质谱法定量。
DOI：
10.1111/irv.12001
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发表时间：
2013-07
影响因子：
4.4
通讯作者：
Alterman M
中科院分区：
文献类型：
--
作者：
Getie-Kebtie M;Sultana I;Eichelberger M;Alterman M
请将本文引用为：Getie-Kebtie 等人。 (2012) 基于无标记质谱法对流感病毒制剂和疫苗中的血凝素和神经氨酸酶进行定量。流感和其他呼吸道病毒 7(4), 521–530。 背景 流感疫苗接种是预防流感及其严重并发症的主要方法。准确快速测定血凝素 (HA) 浓度的方法将有助于参考抗原的制备，从而加快季节性和大流行疫苗的可用性。 目的 本研究的目标是开发一种基于无标记质谱 (MS) 的方法，能够同时识别和定量流感疫苗或病毒制剂中的 HA、神经氨酸酶 (NA) 和其他病毒蛋白和蛋白污染物。 方法 所提出的方法基于对掺有已知量蛋白质标准品的疫苗或病毒制剂胰蛋白酶消化物进行 LC/MSE 分析，从中生成通用响应因子，并应用该因子来计算混合物中鉴定的蛋白质的浓度。 结果 我们表明，通过使用适当的内标，基于无标记 MS 的蛋白质定量方法适用于同时鉴定和绝对定量 HA，以及鉴定和相对定量流感疫苗和病毒制剂中的其他流感蛋白质以及蛋白质杂质。我们表明，不同亚型重组 HA 是首选的内标，可在 HA 和其他流感蛋白的绝对定量方面提供最准确的结果。我们应用这种方法来测量全病毒和单价疫苗制剂中HA的绝对量以及其他病毒蛋白和杂质的相对量，提供数据来证明NA量的毒株依赖性差异。 结论 这里提出的无标记 MS 方法非常适合及时制备季节性和大流行疫苗效力测试所需的参考材料。
Please cite this paper as: Getie‐Kebtie et al. (2012) Label‐free mass spectrometry‐based quantification of hemagglutinin and neuraminidase in influenza virus preparations and vaccines. Influenza and Other Respiratory Viruses 7(4), 521–530. Background Influenza vaccination is the primary method for preventing influenza and its severe complications. An accurate rapid method to determine hemagglutinin (HA) concentration would facilitate reference antigen preparation and consequently expedite availability of seasonal as well as pandemic vaccines. Objective The goal of this study was to develop a label‐free mass spectrometry (MS) based method that enables simultaneous identification and quantification of HA, neuraminidase (NA), and other viral proteins and protein contaminations in influenza vaccine or virus preparations. Methods The method presented is based on LC/MSE analysis of vaccine or virus preparations tryptic digests spiked with a known amount of protein standard from which a universal response factor is generated and applied to calculate the concentration of proteins identified in the mixture. Results We show that, with the use of an appropriate internal standard, the label‐free MS‐based protein quantification method is applicable for simultaneous identification and absolute quantification of HA and identification and relative quantification of other influenza proteins as well as protein impurities in influenza vaccines and virus preparations. We show that different subtype recombinant HA is preferred internal standard that provides the most accurate results in absolute quantification of HAs and other influenza proteins. We applied this method to measure the absolute quantity of HA as well as relative quantities of other viral proteins and impurities in preparations of whole virus and monovalent vaccine, providing data to demonstrate strain‐dependent differences in the amount of NA. Conclusion The label‐free MS method presented here is ideally suited for timely preparation of reference material needed for potency testing of seasonal and pandemic vaccines.
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DOI：
10.1007/978-3-540-92165-3_12
发表时间：
2009-01-01
期刊：
VACCINES FOR PANDEMIC INFLUENZA
影响因子：
--
作者：
Sylte, Matthew J.;Suarez, David L.
通讯作者：
Suarez, David L.
影响因子：
7
作者：
Silva, JC;Gorenstein, MV;Geromanos, SJ
通讯作者：
Geromanos, SJ
影响因子：
5.5
作者：
Farnsworth, Aaron;Cyr, Terry D.;Li, Xuguang
通讯作者：
Li, Xuguang
影响因子：
5.5
作者：
Harvey, Ruth;Wheeler, Jun X.;Engelhardt, Othmar G.
通讯作者：
Engelhardt, Othmar G.
影响因子：
2.9
作者：
Norrgran, Jessica;Williams, Tracie L.;Barr, John R.
通讯作者：
Barr, John R.