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Updated: Oct 21, 2025
Production of a SARS-CoV-2 Virus-Like-Particle System to Investigate Viral Life Cycles In Vitro
Published on: June 6, 2025
Structural characterization of β-propiolactone inactivated severe acute respiratory syndrome coronavirus 2
Dmitry V Bagrov1,2, Grigory S Glukhov1, Andrey V Moiseenko1,3
1Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia.
Abstract:
The severe COVID-19 pandemic drives the research toward the SARS-CoV-2 virion structure and the possible therapies against it. Here, we characterized the β-propiolactone inactivated SARS-CoV-2 virions using transmission electron microscopy (TEM) and atomic force microscopy (AFM). We compared the SARS-CoV-2 samples purified by two consecutive chromatographic procedures (size exclusion chromatography [SEC], followed by ion-exchange chromatography [IEC]) with samples purified by ultracentrifugation. The samples prepared using SEC and IEC retained more spikes on the surface than the ones prepared using ultracentrifugation, as confirmed by TEM and AFM. TEM showed that the spike (S) proteins were in the pre-fusion conformation. Notably, the S proteins could be recognized by specific monoclonal antibodies. Analytical TEM showed that the inactivated virions retained nucleic acid. Altogether, we demonstrated that the inactivated SARS-CoV-2 virions retain the structural features of native viruses and provide a prospective vaccine candidate.
Insights
Inactivated SARS-CoV-2 virions retain key structural features, including surface spikes and nucleic acid, making them potential vaccine candidates. This research aids in developing effective COVID-19 therapies.
Area of Science:
- Virology
- Vaccine Development
- Structural Biology
Background:
- The COVID-19 pandemic necessitates research into SARS-CoV-2 structure and therapies.
- Understanding virion integrity is crucial for vaccine design.
Purpose of the Study:
- To characterize inactivated SARS-CoV-2 virions.
- To compare purification methods for preserving virion structure.
- To evaluate inactivated virions as potential vaccine candidates.
Main Methods:
- Characterization using transmission electron microscopy (TEM) and atomic force microscopy (AFM).
- Comparison of size exclusion chromatography (SEC) with ion-exchange chromatography (IEC) versus ultracentrifugation for purification.
- Analysis of spike protein conformation and antibody recognition.
- Assessment of nucleic acid retention.
Main Results:
- SEC and IEC purification methods retained more surface spikes compared to ultracentrifugation.
- TEM confirmed spike proteins were in a pre-fusion conformation.
- Inactivated virions showed recognition by monoclonal antibodies and retained nucleic acid.
Conclusions:
- β-propiolactone inactivated SARS-CoV-2 virions maintain structural integrity similar to native viruses.
- The structural preservation supports their potential as vaccine candidates.
- Optimized purification methods enhance the suitability of inactivated virions for vaccine development.
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