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High-throughput identification of prefusion-stabilizing mutations in SARS-CoV-2 spike
Timothy J C Tan1, Zongjun Mou2, Ruipeng Lei3
1Center for Biophysics and Quantitative Biology, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
Nature Communications
|April 10, 2023
Summary
A new unbiased method identifies mutations that stabilize the SARS-CoV-2 spike protein for improved COVID-19 vaccine development. This approach enhances protein expression and reduces viral fusion, accelerating immunogen design.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Prefusion-stabilized spike proteins are crucial for effective COVID-19 vaccines.
- Current methods for engineering stabilized spike immunogens are experimentally intensive and rely on structural data.
Purpose of the Study:
- To develop a systematic and unbiased method for identifying mutations that enhance SARS-CoV-2 spike protein expression and stabilize its prefusion conformation.
- To demonstrate the utility of this method in discovering novel stabilizing mutations.
Main Methods:
- Integration of a fluorescence-based fusion assay, mammalian cell display, and deep mutational scanning.
- Application of the method to a specific region within the S2 domain of the SARS-CoV-2 spike protein.
Main Results:
- Identification of mutations, in addition to the known K986P/V987P, that improve spike protein expression.
- Discovery of mutations that significantly reduce spike protein fusogenicity, indicating enhanced prefusion stabilization.
- Validation of a novel, efficient approach for identifying beneficial spike protein mutations.
Conclusions:
- The developed method offers a powerful, unbiased strategy for engineering prefusion-stabilized viral immunogens.
- This approach can accelerate vaccine development not only for SARS-CoV-2 but also for other viral pathogens.
- Findings provide new insights into spike protein stabilization and immunogen design.
