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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-06-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/14/6/1332 |
| _version_ | 1797481368811208704 |
|---|---|
| author | Hancong Sun Jinghan Xu Guanying Zhang Jin Han Meng Hao Zhengshan Chen Ting Fang Xiangyang Chi Changming Yu |
| author_facet | Hancong Sun Jinghan Xu Guanying Zhang Jin Han Meng Hao Zhengshan Chen Ting Fang Xiangyang Chi Changming Yu |
| author_sort | Hancong Sun |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T22:13:35Z |
| format | Article |
| id | doaj.art-a0457817e5d54fd2b337341547c8dd73 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T22:13:35Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-a0457817e5d54fd2b337341547c8dd732023-11-23T19:27:37ZengMDPI AGViruses1999-49152022-06-01146133210.3390/v14061332Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 VariantsHancong Sun0Jinghan Xu1Guanying Zhang2Jin Han3Meng Hao4Zhengshan Chen5Ting Fang6Xiangyang Chi7Changming Yu8Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, ChinaThe 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.https://www.mdpi.com/1999-4915/14/6/1332SARS-CoV-2 variantsOmicronpseudovirusneutralization assayconvalescent plasma |
| spellingShingle | Hancong Sun Jinghan Xu Guanying Zhang Jin Han Meng Hao Zhengshan Chen Ting Fang Xiangyang Chi Changming Yu Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants Viruses SARS-CoV-2 variants Omicron pseudovirus neutralization assay convalescent plasma |
| title | Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants |
| title_full | Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants |
| title_fullStr | Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants |
| title_full_unstemmed | Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants |
| title_short | Developing Pseudovirus-Based Neutralization Assay against Omicron-Included SARS-CoV-2 Variants |
| title_sort | developing pseudovirus based neutralization assay against omicron included sars cov 2 variants |
| topic | SARS-CoV-2 variants Omicron pseudovirus neutralization assay convalescent plasma |
| url | https://www.mdpi.com/1999-4915/14/6/1332 |
| work_keys_str_mv | AT hancongsun developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT jinghanxu developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT guanyingzhang developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT jinhan developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT menghao developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT zhengshanchen developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT tingfang developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT xiangyangchi developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants AT changmingyu developingpseudovirusbasedneutralizationassayagainstomicronincludedsarscov2variants |