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) 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
Microorganisms - 12(2024), 3 vom: 29. Feb. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Cai, Zhaohui [VerfasserIn] |
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| Links: |
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| Themen: |
Assay validation |
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| Anmerkungen: |
Date Revised 30.03.2024 published: Electronic Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3390/microorganisms12030501 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM370330919 |
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| 100 | 1 | |a Cai, Zhaohui |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A Pseudovirus-Based Neutralization Assay for SARS-CoV-2 Variants |b A Rapid, Cost-Effective, BSL-2-Based High-Throughput Assay Useful for Vaccine Immunogenicity Evaluation |
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| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a COVID-19 | |
| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 4 | |a XBB.1.16 | |
| 650 | 4 | |a XBB.1.5 | |
| 650 | 4 | |a XBB.2.3 | |
| 650 | 4 | |a assay validation | |
| 650 | 4 | |a correlate of protection | |
| 650 | 4 | |a immunogenicity | |
| 650 | 4 | |a neutralizing antibody titers | |
| 650 | 4 | |a pseudovirus-based neutralization assays | |
| 700 | 1 | |a Kalkeri, Raj |e verfasserin |4 aut | |
| 700 | 1 | |a Zhu, Mingzhu |e verfasserin |4 aut | |
| 700 | 1 | |a Cloney-Clark, Shane |e verfasserin |4 aut | |
| 700 | 1 | |a Haner, Benjamin |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Mi |e verfasserin |4 aut | |
| 700 | 1 | |a Osman, Bahar |e verfasserin |4 aut | |
| 700 | 1 | |a Dent, Dominic |e verfasserin |4 aut | |
| 700 | 1 | |a Feng, Sheau-Line |e verfasserin |4 aut | |
| 700 | 1 | |a Longacre, Zach |e verfasserin |4 aut | |
| 700 | 1 | |a Glenn, Greg |e verfasserin |4 aut | |
| 700 | 1 | |a Plested, Joyce S |e verfasserin |4 aut | |
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