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Targeted Proteomics for the Detection of SARS-CoV-2 Proteins

Karel Bezstarosti, View ORCID ProfileMart M. Lamers, View ORCID ProfileBart L. Haagmans, View ORCID ProfileJeroen A. A. Demmers
doi: https://doi.org/10.1101/2020.04.23.057810
Karel Bezstarosti
1Proteomics Center, Erasmus University Medical Center, Rotterdam, the Netherlands
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Mart M. Lamers
2Viroscience Department, Erasmus University Medical Center, Rotterdam, the Netherlands
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Bart L. Haagmans
2Viroscience Department, Erasmus University Medical Center, Rotterdam, the Netherlands
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Jeroen A. A. Demmers
1Proteomics Center, Erasmus University Medical Center, Rotterdam, the Netherlands
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  • ORCID record for Jeroen A. A. Demmers
  • For correspondence: j.demmers@erasmusmc.nl
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ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The rapid, sensitive and specific diagnosis of SARS-CoV-2 by fast and unambiguous testing is widely recognized to be critical in responding the current outbreak. Since the current testing capacity by conventional PCR based methods is insufficient because of shortages of supplies such as RNA extraction kits and PCR reagents, alternative and/or complementary testing assays should be developed. Here, we exploit the potential of targeted mass spectrometry based proteomic technologies to solve the current issue of insufficient SARS-CoV-2 diagnostic testing capacity. We have assessed the limit of detection by parallel reaction monitoring (PRM) on an Orbitrap Eclipse mass spectrometer for target tryptic peptides of several SARS-CoV-2 proteins from a sample of virus infected Vero cells. For Nucleocapsid protein the limit of detection was found to be in the mid-attomole range (0.9 × 10−12 g), which would theoretically correspond to approximately 10,000 SARS-CoV-2 particles, under the assumption that all viral proteins are assembled in macromolecular virus particles. Whether or not this sensitivity is sufficient to play a role in SARS-CoV-2 detection in patient material such as swabs or body fluids largely depends on the amount of viral proteins present in such samples and is subject of further research. If yes, mass spectrometry based methods could serve as a complementary protein based diagnostic tool and further steps should be focused on sample preparation protocols and on improvements in sample throughput.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://www.ebi.ac.uk/pride/

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted April 23, 2020.
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Targeted Proteomics for the Detection of SARS-CoV-2 Proteins
Karel Bezstarosti, Mart M. Lamers, Bart L. Haagmans, Jeroen A. A. Demmers
bioRxiv 2020.04.23.057810; doi: https://doi.org/10.1101/2020.04.23.057810
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Targeted Proteomics for the Detection of SARS-CoV-2 Proteins
Karel Bezstarosti, Mart M. Lamers, Bart L. Haagmans, Jeroen A. A. Demmers
bioRxiv 2020.04.23.057810; doi: https://doi.org/10.1101/2020.04.23.057810

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