التفاصيل البيبلوغرافية
| العنوان: |
Inactivation of Coronaviruses during Sample Preparation for Proteomics Experiments |
| المؤلفون: |
Marica Grossegesse (8861096), Paula Leupold (11331716), Joerg Doellinger (294849), Lars Schaade (824526), Andreas Nitsche (12987) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biophysics, Biochemistry, Microbiology, Genetics, Molecular Biology, Neuroscience, Biotechnology, Immunology, Infectious Diseases, Chemical Sciences not elsewhere classified, systematically analyzed yet, 95 ° c, heating step prior, partially incomplete inactivation, choosing lysis buffers, room temperature sds, proteomics lysis buffers, room temperature, step could, proteomics analysis, based proteomics, typical buffers, urea , urea resulted, time pcr, serial passaging, sample preparation, safety reasons, safe inactivation, quantitative real |
| الوصف: |
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 buffers4% SDS, 1% SDC, TFA, 6 M GdmCl, and 8 M ureafor 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. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/journal_contribution/Inactivation_of_Coronaviruses_during_Sample_Preparation_for_Proteomics_Experiments/16441442 |
| DOI: |
10.1021/acs.jproteome.1c00320.s001 |
| الاتاحة: |
https://doi.org/10.1021/acs.jproteome.1c00320.s001 |
| Rights: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.9406383B |
| قاعدة البيانات: |
BASE |