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A Fluorogenic Peptide Cleavage Assay to Screen for Proteolytic Activity: Applications for coronavirus spike protein activation
Published on: January 9, 2019
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Proteases and variants: context matters for SARS-CoV-2 entry assays
Christian S Stevens1, Kasopefoluwa Y Oguntuyo1, Benhur Lee1
1Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, United States.
Current Opinion in Virology
|August 8, 2021
Summary
Understanding SARS-CoV-2 entry complexity is key. Standardizing assays for spike protein activation and cell entry improves research on immunity, therapeutics, and variants.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) utilizes diverse entry mechanisms mediated by its spike (S) protein.
- SARS-CoV-2 entry is contingent upon variable proteolytic priming, activation, and receptor binding, influenced by contextual factors.
Purpose of the Study:
- To review the impact of complex SARS-CoV-2 entry pathways on entry assays.
- To explore how these assays inform our understanding of humoral immunity, therapeutic efficacy, and tissue tropism.
- To highlight the role of spike protein proteolytic activation in understanding viral variants and pathogenicity.
Main Methods:
- Review of existing literature on SARS-CoV-2 entry mechanisms.
- Focus on the proteolytic activation of the SARS-CoV-2 spike protein.
- Analysis of implications for entry assay design and interpretation.
Main Results:
- The complexity of SARS-CoV-2 entry pathways significantly affects the interpretation of entry assays.
- Proteolytic activation of the spike protein is crucial for understanding viral variants, transmission, and pathogenicity.
- Current assay methodologies may lack standardization, impacting reproducibility and comparability.
Conclusions:
- Standardized universal criteria for virus entry assays are needed.
- Best practices for reporting experimental details, including viral passage, cell lines, and proteases, are essential for robust research.
- Improved assay standards will enhance the understanding of SARS-CoV-2 biology and inform therapeutic development.
