Measuring Neutralizing Antibodies to SARS-CoV-2 Using Lentiviral Spike-Pseudoviruses
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature..
Assays measuring neutralizing antibodies (nAbs) against SARS-CoV-2 are used to evaluate serological responses after SARS-CoV-2 infection and the potency of therapeutic antibodies and preventive vaccines. It is therefore imperative that neutralization assays be sensitive, specific, quantitative, and scalable for high throughput. Pseudoviruses are excellent surrogates for highly pathogenic viruses such as SARS-CoV-2 because they can be safely used to measure nAbs in a biosafety level-2 laboratory. In addition, pseudoviruses allow for easy introduction of mutations to study the effect of amino acid changes in the spike protein. In this chapter, we describe a recently optimized assay for measuring neutralizing antibodies to SARS-CoV-2 that uses a HIV-based lentiviral vector pseudotyped with the spike glycoprotein of SARS-CoV-2 to infect 293T cells stably expressing ACE2 and TMPRSS2.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:2452 |
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| Enthalten in: |
Methods in molecular biology (Clifton, N.J.) - 2452(2022) vom: 10., Seite 305-314 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Neerukonda, Sabari Nath [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 17.05.2022 Date Revised 13.06.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.1007/978-1-0716-2111-0_18 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Assays measuring neutralizing antibodies (nAbs) against SARS-CoV-2 are used to evaluate serological responses after SARS-CoV-2 infection and the potency of therapeutic antibodies and preventive vaccines. It is therefore imperative that neutralization assays be sensitive, specific, quantitative, and scalable for high throughput. Pseudoviruses are excellent surrogates for highly pathogenic viruses such as SARS-CoV-2 because they can be safely used to measure nAbs in a biosafety level-2 laboratory. In addition, pseudoviruses allow for easy introduction of mutations to study the effect of amino acid changes in the spike protein. In this chapter, we describe a recently optimized assay for measuring neutralizing antibodies to SARS-CoV-2 that uses a HIV-based lentiviral vector pseudotyped with the spike glycoprotein of SARS-CoV-2 to infect 293T cells stably expressing ACE2 and TMPRSS2 | ||
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| 700 | 1 | |a Wang, Wei |e verfasserin |4 aut | |
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