Department of Mechanical Engineering and Materials Science, Yale University, Mason Laboratory, 9 Hillhouse Avenue, New Haven, Connecticut 06520-8286, United States.
Department of Chemical and Environmental Engineering, Yale University, New Haven, Connecticut 06511, United States.
Anal Chem. 2021 Sep 28;93(38):12938-12943. doi: 10.1021/acs.analchem.1c02402. Epub 2021 Sep 14.
We use the Φ6 bacteriophage previously exploited as a BSL-1 surrogate of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome (MERS) coronavirus to obtain the first high-resolution gas phase mobility spectra of an enveloped virus. The relative full width at half-maximum found for the viral mobility distribution (FWHM < 3.7%) is substantially narrower than that reported by prior mobility or microscopy studies with other enveloped viruses. It is nevertheless not as narrow as that recently found for several non-enveloped viruses (FWHM ≈ 2%), presumably due to particle to particle variability of enveloped viruses. This 3.7% is an upper bound to the actual width. Nevertheless, the well-defined mobility peaks obtained indicate that gas phase mobility analysis is a more discriminating methodology than that previously demonstrated for physically based non-genetic viral diagnostic of enveloped viruses. These results are obtained by analysis of the original cell culture medium containing the virus, purified only by passage through a 0.22 μm filter and by dialysis into a 10 mM aqueous ammonium acetate buffer. We confirmed that this buffer exchange preserves infectivity. Therefore, the 63.7 nm mobility diameter found, although smaller than the 75 nm previously inferred by microscopy, corresponds to the full particle including the envelope.
我们使用先前作为 BSL-1 严重急性呼吸综合征冠状病毒（SARS-CoV）和中东呼吸综合征冠状病毒（MERS-CoV）替代物的 Φ6 噬菌体，获得了首个包膜病毒的高分辨率气相迁移率谱。病毒迁移分布的相对半峰全宽（FWHM < 3.7%）明显比先前用其他包膜病毒进行的迁移率或显微镜研究报告的要窄。然而，它不如最近对几种非包膜病毒（FWHM ≈ 2%）发现的那么窄，这可能是由于包膜病毒的颗粒间变异性所致。这 3.7%是实际宽度的上限。尽管如此，获得的明确迁移峰表明，气相迁移率分析比以前针对包膜病毒的基于物理的非遗传病毒诊断所证明的方法更具辨别力。这些结果是通过对仅通过 0.22 μm 过滤器过滤和通过透析到 10 mM 水性乙酸铵缓冲液中进行纯化的含有病毒的原始细胞培养液进行分析得到的。我们证实，这种缓冲液交换可以保持感染力。因此，尽管发现的 63.7nm 迁移直径小于以前通过显微镜推断的 75nm，但它对应于包括包膜的完整颗粒。
