Preparation and characterization of inactivated tick-borne encephalitis virus samples for single-particle imaging at the European XFEL.

Acta crystallographica. Section D, Structural biology 2024 Jan 01

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X-ray imaging of virus particles at the European XFEL could eventually allow their complete structures to be solved, potentially approaching the resolution of other structural virology methods. To achieve this ambitious goal with today's technologies, about 1 ml of purified virus suspension containing at least 1012 particles per millilitre is required. Such large amounts of concentrated suspension have never before been obtained for enveloped viruses. Tick-borne encephalitis virus (TBEV) represents an attractive model system for the development of enveloped virus purification and concentration protocols, given the availability of large amounts of inactivated virus material provided by vaccine-manufacturing facilities. Here, the development of a TBEV vaccine purification and concentration scheme is presented combined with a quality-control protocol that allows substantial amounts of highly concentrated non-aggregated suspension to be obtained. Preliminary single-particle imaging experiments were performed for this sample at the European XFEL, showing distinct diffraction patterns.

Citation

Mikhail F Vorovitch, Valeriya R Samygina, Evgeny Pichkur, Peter V Konarev, Georgy Peters, Evgeny V Khvatov, Alla L Ivanova, Ksenia K Tuchynskaya, Olga I Konyushko, Anton Y Fedotov, Grigory Armeev, Konstantin V Shaytan, Mikhail V Kovalchuk, Dmitry I Osolodkin, Alexey M Egorov, Aydar A Ishmukhametov. Preparation and characterization of inactivated tick-borne encephalitis virus samples for single-particle imaging at the European XFEL. Acta crystallographica. Section D, Structural biology. 2024 Jan 01;80(Pt 1):44-59