Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona, 7410001, Israel.
Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona, 7410001, Israel.
J Virol Methods. 2022 May;303:114498. doi: 10.1016/j.jviromet.2022.114498. Epub 2022 Feb 22.
The spike glycoprotein mediates virus binding to the host cells and is a key target for vaccines development. One SARS-CoV-2 vaccine is based on vesicular stomatitis virus (VSV), in which the native surface glycoprotein has been replaced by the SARS-CoV-2 spike protein (VSV-ΔG-spike). The titer of the virus is quantified by the plaque forming unit (PFU) assay, but there is no method for spike protein quantitation as an antigen in a VSV-based vaccine. Here, we describe a mass spectrometric (MS) spike protein quantification method, applied to VSV-ΔG-spike based vaccine. Proof of concept of this method, combining two different sample preparations, is shown for complex matrix samples, produced during the vaccine manufacturing processes. Total spike levels were correlated with results from activity assays, and ranged between 0.3-0.5 μg of spike protein per 10 PFU virus-based vaccine. This method is simple, linear over a wide range, allows quantification of antigen within a sample and can be easily implemented for any vaccine or therapeutic sample.
刺突糖蛋白介导病毒与宿主细胞的结合，是疫苗开发的关键靶点。一种基于水疱性口炎病毒（VSV）的 SARS-CoV-2 疫苗，其中已用 SARS-CoV-2 刺突蛋白（VSV-ΔG-spike）取代了天然表面糖蛋白。病毒的效价通过蚀斑形成单位（PFU）测定法进行定量，但作为基于 VSV 的疫苗中的抗原，没有针对刺突蛋白定量的方法。在这里，我们描述了一种用于 VSV-ΔG-spike 基于疫苗的质谱（MS）刺突蛋白定量方法。该方法的概念验证，结合了两种不同的样品制备方法，用于疫苗生产过程中产生的复杂基质样品。总刺突水平与活性测定结果相关，范围在每 10 PFU 病毒疫苗 0.3-0.5 μg 的刺突蛋白之间。该方法简单，线性范围宽，允许在样品内定量抗原，并且可以很容易地应用于任何疫苗或治疗性样品。
