Tags

Type your tag names separated by a space and hit enter

Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure.
Viruses. 2021 03 26; 13(4)V

Abstract

The risk posed by Severe Acute Respiratory Syndrome Coronavirus -2 (SARS-CoV-2) dictates that live-virus research is conducted in a biosafety level 3 (BSL3) facility. Working with SARS-CoV-2 at lower biosafety levels can expedite research yet requires the virus to be fully inactivated. In this study, we validated and compared two protocols for inactivating SARS-CoV-2: heat treatment and ultraviolet irradiation. The two methods were optimized to render the virus completely incapable of infection while limiting the destructive effects of inactivation. We observed that 15 min of incubation at 65 °C completely inactivates high titer viral stocks. Complete inactivation was also achieved with minimal amounts of UV power (70,000 µJ/cm2), which is 100-fold less power than comparable studies. Once validated, the two methods were then compared for viral RNA quantification, virion purification, and antibody detection assays. We observed that UV irradiation resulted in a 2-log reduction of detectable genomes compared to heat inactivation. Protein yield following virion enrichment was equivalent for all inactivation conditions, but the quality of resulting viral proteins and virions were differentially impacted depending on inactivation method and time. Here, we outline the strengths and weaknesses of each method so that investigators might choose the one which best meets their research goals.

Authors+Show Affiliations

Department of Chemical & Biological Engineering, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Plant Science and Plant Pathology, Montana State University; Bozeman, MT 59717, USA.Department of Plant Science and Plant Pathology, Montana State University; Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.Department of Chemical & Biological Engineering, Montana State University, Bozeman, MT 59717, USA.Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA.

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33810401

Citation

Loveday, Emma K., et al. "Effect of Inactivation Methods On SARS-CoV-2 Virion Protein and Structure." Viruses, vol. 13, no. 4, 2021.
Loveday EK, Hain KS, Kochetkova I, et al. Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure. Viruses. 2021;13(4).
Loveday, E. K., Hain, K. S., Kochetkova, I., Hedges, J. F., Robison, A., Snyder, D. T., Brumfield, S. K., Young, M. J., Jutila, M. A., Chang, C. B., & Taylor, M. P. (2021). Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure. Viruses, 13(4). https://doi.org/10.3390/v13040562
Loveday EK, et al. Effect of Inactivation Methods On SARS-CoV-2 Virion Protein and Structure. Viruses. 2021 03 26;13(4) PubMed PMID: 33810401.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure. AU - Loveday,Emma K, AU - Hain,Kyle S, AU - Kochetkova,Irina, AU - Hedges,Jodi F, AU - Robison,Amanda, AU - Snyder,Deann T, AU - Brumfield,Susan K, AU - Young,Mark J, AU - Jutila,Mark A, AU - Chang,Connie B, AU - Taylor,Matthew P, Y1 - 2021/03/26/ PY - 2021/03/09/received PY - 2021/03/22/accepted PY - 2021/4/3/entrez PY - 2021/4/4/pubmed PY - 2021/4/17/medline KW - ELISA KW - SARS-CoV-2 KW - UV inactivation KW - electron microscopy KW - heat inactivation KW - western blot JF - Viruses JO - Viruses VL - 13 IS - 4 N2 - The risk posed by Severe Acute Respiratory Syndrome Coronavirus -2 (SARS-CoV-2) dictates that live-virus research is conducted in a biosafety level 3 (BSL3) facility. Working with SARS-CoV-2 at lower biosafety levels can expedite research yet requires the virus to be fully inactivated. In this study, we validated and compared two protocols for inactivating SARS-CoV-2: heat treatment and ultraviolet irradiation. The two methods were optimized to render the virus completely incapable of infection while limiting the destructive effects of inactivation. We observed that 15 min of incubation at 65 °C completely inactivates high titer viral stocks. Complete inactivation was also achieved with minimal amounts of UV power (70,000 µJ/cm2), which is 100-fold less power than comparable studies. Once validated, the two methods were then compared for viral RNA quantification, virion purification, and antibody detection assays. We observed that UV irradiation resulted in a 2-log reduction of detectable genomes compared to heat inactivation. Protein yield following virion enrichment was equivalent for all inactivation conditions, but the quality of resulting viral proteins and virions were differentially impacted depending on inactivation method and time. Here, we outline the strengths and weaknesses of each method so that investigators might choose the one which best meets their research goals. SN - 1999-4915 UR - https://www.unboundmedicine.com/medline/citation/33810401/Effect_of_Inactivation_Methods_on_SARS_CoV_2_Virion_Protein_and_Structure_ L2 - https://www.mdpi.com/resolver?pii=v13040562 DB - PRIME DP - Unbound Medicine ER -