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Article

Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants

Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China
*
Authors to whom correspondence should be addressed.
Academic Editors: Ahmed Elshamy and Mohamed Ibrahim
Viruses 2022, 14(6), 1332; https://doi.org/10.3390/v14061332
Received: 5 April 2022 / Revised: 6 June 2022 / Accepted: 16 June 2022 / Published: 18 June 2022
The global spread of SARS-CoV-2 and its variants poses a serious threat to human health worldwide. Recently, the emergence of Omicron has presented a new challenge to the prevention and control of the COVID-19 pandemic. A convenient and reliable in vitro neutralization assay is an important method for validating the efficiency of antibodies, vaccines, and other potential drugs. Here, we established an effective assay based on a pseudovirus carrying a full-length spike (S) protein of SARS-CoV-2 variants in the HIV-1 backbone, with a luciferase reporter gene inserted into the non-replicate pseudovirus genome. The key parameters for packaging the pseudovirus were optimized, including the ratio of the S protein expression plasmids to the HIV backbone plasmids and the collection time for the Alpha, Beta, Gamma, Kappa, and Omicron pseudovirus particles. The pseudovirus neutralization assay was validated using several approved or developed monoclonal antibodies, underscoring that Omicron can escape some neutralizing antibodies, such as REGN10987 and REGN10933, while S309 and ADG-2 still function with reduced neutralization capability. The neutralizing capacity of convalescent plasma from COVID-19 convalescent patients in Wuhan was tested against these pseudoviruses, revealing the immune evasion of Omicron. Our work established a practical pseudovirus-based neutralization assay for SARS-CoV-2 variants, which can be conducted safely under biosafety level-2 (BSL-2) conditions, and this assay will be a promising tool for studying and characterizing vaccines and therapeutic candidates against Omicron-included SARS-CoV-2 variants. View Full-Text
Keywords: SARS-CoV-2 variants; Omicron; pseudovirus; neutralization assay; convalescent plasma SARS-CoV-2 variants; Omicron; pseudovirus; neutralization assay; convalescent plasma
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MDPI and ACS Style

Sun, H.; Xu, J.; Zhang, G.; Han, J.; Hao, M.; Chen, Z.; Fang, T.; Chi, X.; Yu, C. Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants. Viruses 2022, 14, 1332. https://doi.org/10.3390/v14061332

AMA Style

Sun H, Xu J, Zhang G, Han J, Hao M, Chen Z, Fang T, Chi X, Yu C. Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants. Viruses. 2022; 14(6):1332. https://doi.org/10.3390/v14061332

Chicago/Turabian Style

Sun, Hancong, Jinghan Xu, Guanying Zhang, Jin Han, Meng Hao, Zhengshan Chen, Ting Fang, Xiangyang Chi, and Changming Yu. 2022. "Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants" Viruses 14, no. 6: 1332. https://doi.org/10.3390/v14061332

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