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真核无细胞系统作为对新型人畜共患病原体快速反应的潜力:SARS-CoV-2病毒蛋白分析

The Potential of Eukaryotic Cell-Free Systems as a Rapid Response to Novel Zoonotic Pathogens: Analysis of SARS-CoV-2 Viral Proteins.

作者信息

Ramm Franziska, Dondapati Srujan K, Trinh Hoai Anh, Wenzel Dana, Walter Ruben M, Zemella Anne, Kubick Stefan

机构信息

Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), Potsdam, Germany.

Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany.

出版信息

Front Bioeng Biotechnol. 2022 Apr 19;10:896751. doi: 10.3389/fbioe.2022.896751. eCollection 2022.

Abstract

The ongoing pandemic caused by the novel coronavirus (SARS-CoV-2) has led to more than 445 million infections and the underlying disease, COVID-19, resulted in more than 6 million deaths worldwide. The scientific world is already predicting future zoonotic diseases. Hence, rapid response systems are needed to tackle future epidemics and pandemics. Here, we present the use of eukaryotic cell-free systems for the rapid response to novel zoonotic diseases represented by SARS-CoV-2. Non-structural, structural and accessory proteins encoded by SARS-CoV-2 were synthesized by cell-free protein synthesis in a fast and efficient manner. The inhibitory effect of the non-structural protein 1 on protein synthesis could be shown . Structural proteins were quantitatively detected by commercial antibodies, therefore facilitating cell-free systems for the validation of available antibodies. The cytotoxic envelope protein was characterized in electrophysiological planar lipid bilayer measurements. Hence, our study demonstrates the potential of eukaryotic cell-free systems as a rapid response mechanism for the synthesis, functional characterization and antibody validation against a viral pathogen.

摘要

新型冠状病毒(SARS-CoV-2)引发的持续大流行已导致超过4.45亿人感染,其所致的基础疾病——新冠肺炎,在全球范围内造成了超过600万人死亡。科学界已经在预测未来的人畜共患疾病。因此,需要快速反应系统来应对未来的流行病和大流行。在此,我们展示了真核无细胞系统在快速应对以SARS-CoV-2为代表的新型人畜共患疾病方面的应用。SARS-CoV-2编码的非结构蛋白、结构蛋白和辅助蛋白通过无细胞蛋白质合成以快速高效的方式合成。可以证明非结构蛋白1对蛋白质合成的抑制作用。结构蛋白通过商用抗体进行定量检测,从而便于利用无细胞系统验证现有抗体。在电生理平面脂质双层测量中对具有细胞毒性的包膜蛋白进行了表征。因此,我们的研究证明了真核无细胞系统作为一种快速反应机制在针对病毒病原体的合成、功能表征和抗体验证方面的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07cb/9061942/fa3652e00eba/fbioe-10-896751-g001.jpg

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