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Inactivation of Coronaviruses during Sample Preparation for Proteomics Experiments.
Grossegesse, Marica; Leupold, Paula; Doellinger, Joerg; Schaade, Lars; Nitsche, Andreas.
  • Grossegesse M; Robert Koch Institute, Centre for Biological Threats and Special Pathogens, ZBS 1: Highly Pathogenic Viruses, Seestr. 10, 13353, Berlin, Germany.
  • Leupold P; Robert Koch Institute, Centre for Biological Threats and Special Pathogens, ZBS 1: Highly Pathogenic Viruses, Seestr. 10, 13353, Berlin, Germany.
  • Doellinger J; Robert Koch Institute, Centre for Biological Threats and Special Pathogens, ZBS 1: Highly Pathogenic Viruses, Seestr. 10, 13353, Berlin, Germany.
  • Schaade L; Robert Koch Institute, Centre for Biological Threats and Special Pathogens, ZBS 6: Proteomics and Spectroscopy, Seestr. 10, 13353, Berlin, Germany.
  • Nitsche A; Robert Koch Institute, Centre for Biological Threats and Special Pathogens, Seestr. 10, 13353, Berlin, Germany.
J Proteome Res ; 20(9): 4598-4602, 2021 09 03.
Article in English | MEDLINE | ID: covidwho-1371586
ABSTRACT
Mass spectrometry-based proteomics is applied in SARS-CoV-2 research and is, moreover, being discussed as a novel method for SARS-CoV-2 diagnostics. However, the safe inactivation of coronaviruses by proteomics lysis buffers has not been systematically analyzed yet. Hence, for safety reasons a heating step prior to sample preparation is often performed. This step could be omitted once the safe inactivation with the typical buffers is proven. Here we test five different proteomics lysis buffers-4% SDS, 1% SDC, TFA, 6 M GdmCl, and 8 M urea-for their inactivation capacity of coronaviruses. Two representative human coronaviruses, namely HCoV-229E and HCoV-OC43, were used as surrogate for SARS-CoV-2. Lysis was performed at room temperature and at 95 °C for 5 min. Inactivation was confirmed by the absence of a cytopathic effect in MRC-5 cells, and equivocal results were further confirmed by serial passaging and quantitative real-time PCR. While at room temperature SDS, SDC, and TFA inactivated both coronaviruses, and GdmCl and urea resulted in partially incomplete inactivation. This demonstrates that care should be taken when choosing lysis buffers for proteomics analysis of coronaviruses, because some buffers do not ensure inactivation and, hence, biosafety during the further sample preparation.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Coronavirus 229E, Human / Coronavirus OC43, Human / COVID-19 Limits: Humans Language: English Journal: J Proteome Res Journal subject: Biochemistry Year: 2021 Document Type: Article Affiliation country: Acs.jproteome.1c00320

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Coronavirus 229E, Human / Coronavirus OC43, Human / COVID-19 Limits: Humans Language: English Journal: J Proteome Res Journal subject: Biochemistry Year: 2021 Document Type: Article Affiliation country: Acs.jproteome.1c00320