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Preliminary Structural Study of Inactivated Yellow Fever Virus

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Abstract

Here we present a protocol for the isolation, purification, and inactivation of the yellow fever virus (YFV) aimed to facilitate high-resolution structure determination using cryo-electron microscopy (cryo-EM). Cryo-EM studies require the concentration of viral particles to be at least 1012 particle/mL. The concentration of the sample was evaluated by a combination of methods, including the plaque-forming assay, spectrophotometry, and protein polyacrylamide gel electrophoresis. High purity of the sample was confirmed by electrophoresis and transmission electron microscopy. The cryo-EM data set was collected and the 2D classification of viral particles was performed.

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ACKNOWLEDGMENTS

We are grateful to K.K. Tuchinskaya for discussion and valuable advice.

Funding

The study was supported by the Russian Foundation for Basic Research (project no. 18-02-40026).

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Authors and Affiliations

  1. National Research Centre “Kurchatov Institute”, 123182, Moscow, Russia

    E. B. Pichkur & V. R. Samygina

  2. Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics,” Russian Academy of Sciences, 119333, Moscow, Russia

    E. B. Pichkur & V. R. Samygina

  3. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products, Russian Academy of Sciences, 108819, Moscow, Russia

    A. L. Ivanova, A. Yu. Fedotov, A. P. Ivanov, E. V. Khvatov, A. A. Ishmukhametov & M. F. Vorovich

  4. Sechenov First Moscow State Medical University, 119991, Moscow, Russia

    A. A. Ishmukhametov & M. F. Vorovich

Authors
  1. E. B. Pichkur
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  2. V. R. Samygina
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  3. A. L. Ivanova
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  4. A. Yu. Fedotov
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  5. A. P. Ivanov
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  6. E. V. Khvatov
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  7. A. A. Ishmukhametov
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  8. M. F. Vorovich
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Corresponding author

Correspondence to V. R. Samygina.

Additional information

Translated by T. Safonova

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Pichkur, E.B., Samygina, V.R., Ivanova, A.L. et al. Preliminary Structural Study of Inactivated Yellow Fever Virus. Crystallogr. Rep. 65, 915–921 (2020). https://doi.org/10.1134/S1063774520060267

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  • DOI: https://doi.org/10.1134/S1063774520060267

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