A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation
Neutralizing antibody responses from COVID-19 vaccines are pivotal in conferring protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Effective COVID-19 vaccines and assays measuring neutralizing antibodies against emerging variants (i.e., XBB.1.5, XBB.1.16, and XBB.2.3)...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/12/3/501 |
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| author | Zhaohui Cai Raj Kalkeri Mingzhu Zhu Shane Cloney-Clark Benjamin Haner Mi Wang Bahar Osman Dominic Dent Sheau-Line Feng Zach Longacre Greg Glenn Joyce S. Plested |
| author_facet | Zhaohui Cai Raj Kalkeri Mingzhu Zhu Shane Cloney-Clark Benjamin Haner Mi Wang Bahar Osman Dominic Dent Sheau-Line Feng Zach Longacre Greg Glenn Joyce S. Plested |
| author_sort | Zhaohui Cai |
| collection | DOAJ |
| description | Neutralizing antibody responses from COVID-19 vaccines are pivotal in conferring protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Effective COVID-19 vaccines and assays measuring neutralizing antibodies against emerging variants (i.e., XBB.1.5, XBB.1.16, and XBB.2.3) are needed. The use of biosafety level (BSL)-3 laboratories for live virus assays results in higher costs and a longer turnaround time; therefore, a BSL-2–based pseudovirus neutralization assay (PNT) was developed. The pseudoviruses were produced by cotransfecting cells with plasmids encoding a lentiviral backbone-expressing luciferase reporter; non-surface proteins for lentiviral production; and ancestral or Omicron (BA.1 and BA.5) SARS-CoV-2 spike (S) proteins. The PNT was developed and optimized in dose and kinetics experiments. The representative serum samples (COVID-19–convalescent or NVX-CoV2373–vaccinated participants enrolled in the 2019nCoV-101 trial) demonstrated a wide dynamic range. The neutralization data showed robust correlation with validated anti-recombinant spike IgG levels and angiotensin-converting enzyme 2 inhibition titers (ancestral). This assay is suitable for measurement of the neutralization ability in clinical samples from individuals infected with SARS-CoV-2 or immunized with a COVID-19 vaccine. The results suggest that this PNT provides a lower cost, high-throughput, rapid turnaround alternative to BSL-3–based microneutralization assays and enables the discovery and development of effective vaccines against emerging variants. |
| first_indexed | 2024-04-24T18:00:24Z |
| format | Article |
| id | doaj.art-5858950851b847d3a887997ce88e0ecc |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-04-24T18:00:24Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-5858950851b847d3a887997ce88e0ecc2024-03-27T13:55:39ZengMDPI AGMicroorganisms2076-26072024-02-0112350110.3390/microorganisms12030501A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity EvaluationZhaohui Cai0Raj Kalkeri1Mingzhu Zhu2Shane Cloney-Clark3Benjamin Haner4Mi Wang5Bahar Osman6Dominic Dent7Sheau-Line Feng8Zach Longacre9Greg Glenn10Joyce S. Plested11Clinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USADiscovery, Novavax, Gaithersburg, MD 20878, USADiscovery, Novavax, Gaithersburg, MD 20878, USAClinical Immunology, Novavax, Gaithersburg, MD 20878, USANeutralizing antibody responses from COVID-19 vaccines are pivotal in conferring protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Effective COVID-19 vaccines and assays measuring neutralizing antibodies against emerging variants (i.e., XBB.1.5, XBB.1.16, and XBB.2.3) are needed. The use of biosafety level (BSL)-3 laboratories for live virus assays results in higher costs and a longer turnaround time; therefore, a BSL-2–based pseudovirus neutralization assay (PNT) was developed. The pseudoviruses were produced by cotransfecting cells with plasmids encoding a lentiviral backbone-expressing luciferase reporter; non-surface proteins for lentiviral production; and ancestral or Omicron (BA.1 and BA.5) SARS-CoV-2 spike (S) proteins. The PNT was developed and optimized in dose and kinetics experiments. The representative serum samples (COVID-19–convalescent or NVX-CoV2373–vaccinated participants enrolled in the 2019nCoV-101 trial) demonstrated a wide dynamic range. The neutralization data showed robust correlation with validated anti-recombinant spike IgG levels and angiotensin-converting enzyme 2 inhibition titers (ancestral). This assay is suitable for measurement of the neutralization ability in clinical samples from individuals infected with SARS-CoV-2 or immunized with a COVID-19 vaccine. The results suggest that this PNT provides a lower cost, high-throughput, rapid turnaround alternative to BSL-3–based microneutralization assays and enables the discovery and development of effective vaccines against emerging variants.https://www.mdpi.com/2076-2607/12/3/501COVID-19pseudovirus-based neutralization assaysimmunogenicitySARS-CoV-2assay validationneutralizing antibody titers |
| spellingShingle | Zhaohui Cai Raj Kalkeri Mingzhu Zhu Shane Cloney-Clark Benjamin Haner Mi Wang Bahar Osman Dominic Dent Sheau-Line Feng Zach Longacre Greg Glenn Joyce S. Plested A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation Microorganisms COVID-19 pseudovirus-based neutralization assays immunogenicity SARS-CoV-2 assay validation neutralizing antibody titers |
| title | A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
| title_full | A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
| title_fullStr | A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
| title_full_unstemmed | A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
| title_short | A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants: A Rapid, Cost-Effective, BSL-2–Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
| title_sort | pseudovirus based neutralization assay for sars cov 2 variants a rapid cost effective bsl 2 based high throughput assay useful for vaccine immunogenicity evaluation |
| topic | COVID-19 pseudovirus-based neutralization assays immunogenicity SARS-CoV-2 assay validation neutralizing antibody titers |
| url | https://www.mdpi.com/2076-2607/12/3/501 |
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