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

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 f...

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Bibliographic Details
Published in:Viruses 2022-06, Vol.14 (6), p.1332
Main Authors: Sun, Hancong, Xu, Jinghan, Zhang, Guanying, Han, Jin, Hao, Meng, Chen, Zhengshan, Fang, Ting, Chi, Xiangyang, Yu, Changming
Format: Article
Language:English
Subjects:
Antibodies
Cloning
convalescent plasma
Coronaviruses
COVID-19
COVID-19 vaccines
Genes
Genomes
HIV
Human immunodeficiency virus
Immunosuppressive agents
Infections
Laboratories
Monoclonal antibodies
Mutation
neutralization assay
Omicron
Plasmids
Protein expression
Proteins
pseudovirus
Reporter gene
SARS-CoV-2 variants
Severe acute respiratory syndrome coronavirus 2
Vaccines
Viruses
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Summary: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.
ISSN:1999-4915
1999-4915
DOI:10.3390/v14061332