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Preparation and characterization of inactivated tick-borne encephalitis virus samples for single-particle imaging at the European XFEL
Acta Crystallographica Section D ( IF 2.6 ) Pub Date : 2024-01-02 , DOI: 10.1107/s2059798323010562 Mikhail F Vorovitch 1 , Valeriya R Samygina 2 , Evgeny Pichkur 2 , Peter V Konarev 2 , Georgy Peters 2 , Evgeny V Khvatov 3 , Alla L Ivanova 3 , Ksenia K Tuchynskaya 3 , Olga I Konyushko 3 , Anton Y Fedotov 3 , Grigory Armeev 4 , Konstantin V Shaytan 4 , Mikhail V Kovalchuk 2 , Dmitry I Osolodkin 3 , Alexey M Egorov 3 , Aydar A Ishmukhametov 3
Acta Crystallographica Section D ( IF 2.6 ) Pub Date : 2024-01-02 , DOI: 10.1107/s2059798323010562 Mikhail F Vorovitch 1 , Valeriya R Samygina 2 , Evgeny Pichkur 2 , Peter V Konarev 2 , Georgy Peters 2 , Evgeny V Khvatov 3 , Alla L Ivanova 3 , Ksenia K Tuchynskaya 3 , Olga I Konyushko 3 , Anton Y Fedotov 3 , Grigory Armeev 4 , Konstantin V Shaytan 4 , Mikhail V Kovalchuk 2 , Dmitry I Osolodkin 3 , Alexey M Egorov 3 , Aydar A Ishmukhametov 3
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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.
中文翻译：
在欧洲 XFEL 上进行单颗粒成像灭活蜱传脑炎病毒样品的制备和表征
欧洲 XFEL 对病毒颗粒进行 X 射线成像最终可以解析其完整结构，有可能接近其他结构病毒学方法的分辨率。为了利用当今的技术实现这一雄心勃勃的目标，需要大约1毫升每毫升至少含有10 12 个颗粒的纯化病毒悬浮液。对于有包膜病毒来说，以前从未获得过如此大量的浓缩悬浮液。鉴于疫苗生产设施提供了大量灭活病毒材料，蜱传脑炎病毒（TBEV）代表了一种用于开发包膜病毒纯化和浓缩方案的有吸引力的模型系统。在此，结合质量控制方案提出了 TBEV 疫苗纯化和浓缩方案的开发，该方案允许获得大量高度浓缩的非聚集悬浮液。在欧洲 XFEL 上对该样品进行了初步的单粒子成像实验，显示出不同的衍射图案。
更新日期：2024-01-02
中文翻译：
在欧洲 XFEL 上进行单颗粒成像灭活蜱传脑炎病毒样品的制备和表征
欧洲 XFEL 对病毒颗粒进行 X 射线成像最终可以解析其完整结构，有可能接近其他结构病毒学方法的分辨率。为了利用当今的技术实现这一雄心勃勃的目标，需要大约1毫升每毫升至少含有10 12 个颗粒的纯化病毒悬浮液。对于有包膜病毒来说，以前从未获得过如此大量的浓缩悬浮液。鉴于疫苗生产设施提供了大量灭活病毒材料，蜱传脑炎病毒（TBEV）代表了一种用于开发包膜病毒纯化和浓缩方案的有吸引力的模型系统。在此，结合质量控制方案提出了 TBEV 疫苗纯化和浓缩方案的开发，该方案允许获得大量高度浓缩的非聚集悬浮液。在欧洲 XFEL 上对该样品进行了初步的单粒子成像实验，显示出不同的衍射图案。
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