Characterization of cells susceptible to SARS-COV-2 and methods for detection of neutralizing antibody by focus forming assay
Abstract The SARS-CoV-2 outbreak and subsequent COVID-19 pandemic have highlighted the urgent need to determine what cells are susceptible to infection and for assays to detect and quantify SARS-CoV-2. Furthermore, the ongoing efforts for vaccine development have necessitated the development of rapid, high-throughput methods of quantifying infectious SARS-CoV-2, as well as the ability to screen human polyclonal sera samples for neutralizing antibodies against SARS-CoV-2. To this end, our lab has adapted focus forming assays for SARS-CoV-2 using Vero CCL-81 cells, referred to in this text as Vero WHO. Using the focus forming assay as the basis for screening cell susceptibility and to develop a focus reduction neutralization test. We have shown that this assay is a sensitive tool for determining SARS-CoV-2 neutralizing antibody titer in human, non-human primate, and mouse polyclonal sera following SARS-CoV-2 exposure. Additionally, we describe the viral growth kinetics of SARS-CoV-2 in a variety of different immortalized cell lines and demonstrate via human ACE2 and viral spike protein expression that these cell lines can support viral entry and replication..
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Preprint| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 06. Okt. Zur Gesamtaufnahme - year:2023 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Stone, E. Taylor [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2020.08.20.259838 |
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| PPN (Katalog-ID): |
XBI01862801X |
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| 245 | 1 | 0 | |a Characterization of cells susceptible to SARS-COV-2 and methods for detection of neutralizing antibody by focus forming assay |
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| 520 | |a Abstract The SARS-CoV-2 outbreak and subsequent COVID-19 pandemic have highlighted the urgent need to determine what cells are susceptible to infection and for assays to detect and quantify SARS-CoV-2. Furthermore, the ongoing efforts for vaccine development have necessitated the development of rapid, high-throughput methods of quantifying infectious SARS-CoV-2, as well as the ability to screen human polyclonal sera samples for neutralizing antibodies against SARS-CoV-2. To this end, our lab has adapted focus forming assays for SARS-CoV-2 using Vero CCL-81 cells, referred to in this text as Vero WHO. Using the focus forming assay as the basis for screening cell susceptibility and to develop a focus reduction neutralization test. We have shown that this assay is a sensitive tool for determining SARS-CoV-2 neutralizing antibody titer in human, non-human primate, and mouse polyclonal sera following SARS-CoV-2 exposure. Additionally, we describe the viral growth kinetics of SARS-CoV-2 in a variety of different immortalized cell lines and demonstrate via human ACE2 and viral spike protein expression that these cell lines can support viral entry and replication. | ||
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| 700 | 1 | |a Geerling, Elizabeth |0 (orcid)0000-0002-5104-499X |4 aut | |
| 700 | 1 | |a Steffen, Tara L. |0 (orcid)0000-0002-4569-315X |4 aut | |
| 700 | 1 | |a Hassert, Mariah |0 (orcid)0000-0002-2141-3344 |4 aut | |
| 700 | 1 | |a Dickson, Alexandria |0 (orcid)0000-0002-2282-9069 |4 aut | |
| 700 | 1 | |a Spencer, Jacqueline F. |0 (orcid)0000-0002-9568-4374 |4 aut | |
| 700 | 1 | |a Toth, Karoly |0 (orcid)0000-0001-6624-9683 |4 aut | |
| 700 | 1 | |a DiPaolo, Richard J. |0 (orcid)0000-0002-2191-6689 |4 aut | |
| 700 | 1 | |a Brien, James D. |0 (orcid)0000-0002-1670-8041 |4 aut | |
| 700 | 1 | |a Pinto, Amelia K. |0 (orcid)0000-0002-7147-3978 |4 aut | |
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