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bioRxiv · 10.1101/2024.05.24.595851

Cell-free synthesis and purification of recombinant nucleocapsid (N), membrane (M), and envelope (E) proteins

Wang, L.·Fei, M.·Zhang, W.·Lin, S.·Jiang, Z.·Zhang, S.·Wang, Y.
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Abstract

Rapid production of soluble recombinant antigens is important for developing in vitro diagnostics, vaccines, and drugs against virus such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this research, hard-to-express nucleocapsid, membrane, and envelope proteins were successfully expressed by an Escherichia coli-based cell-free protein synthesis system. The solubility of the proteins was optimized using various amphipathic molecules. Most of the impurities were easily removed by a one-step Ni-NTA affinity chromatography. This study provides an easy and quick alternative for viruss trans-membrane and nucleotides associated recombinant protein expression, which has potential downstream application for early screening of newly emerging viruses.

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Wang, L., Fei, M., Zhang, W., Lin, S., Jiang, Z., Zhang, S., Wang, Y.. 2024-05-27. Cell-free synthesis and purification of recombinant nucleocapsid (N), membrane (M), and envelope (E) proteins. https://doi.org/10.1101/2024.05.24.595851

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