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2D NMR spectra of SARS-CoV-2 proteins.

As part of the AMED project "Project for Developing Technologies for Addressing the Novel Coronavirus Disease and Other Infectious Diseases “, Life science and Biotechnology Section in AIST is driving a project to aim "Development of a Reliable Rapid Diagnostic System for SARS-CoV-2" (Representative: Kimihiko Kameyama). For this purpose, obtaining high quality viral antigens with structural homogeneity is required. The Dynamic Pharmaco-Modalities Research Group and the Biological Data Science Group in the Cellular and Molecular Bioengineering Research Institute have successfully expressed and purified 27 of the 45 structural domains of the SARS-CoV-2 proteins predicted by sequence analysis, using a cell-free expression system. These include functionality important membrane proteins and extracellular proteins and corresponds to more than 50% of ORF in the virus. We also succeeded in synthesizing and purifying atypical sequences such as ORF3b and ORF9b, which have been pointed out to have bioactivity. In addition, 2D NMR spectra of 13 structural domains, including one atypical sequence were obtained. The NMR spectra not only ensure the structural homogeneity and quality of the antigen, but also enable us to analyze the interaction and structure of the SARS-CoV-2 proteins at atomic level, which will be useful for drug developments and basic biological research.

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2D NMR of SARS-CoV-2 proteins obtained from cell-free expression system.

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