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Qualification of ELISA and neutralization methodologies to measure SARS-CoV-2 humoral immunity using human clinical samples

bioRxiv, 2021-07

2021. This article is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (“the License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. ;DOI: 10.1101/2021.07.02.450915

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Title:
Qualification of ELISA and neutralization methodologies to measure SARS-CoV-2 humoral immunity using human clinical samples
Author: Larsen, Sasha E ; Berube, Bryan J ; Pecor, Tiffany ; Cross, Evan ; Brown, Bryan Paul ; Williams, Brittany ; Johnson, Emma ; Qu, Pingping ; Carter, Lauren ; Wren, Samuel ; Kepl, Elizabeth ; Sydeman, Claire ; King, Neil P ; Baldwin, Susan L ; Coler, Rhea N
Subjects: Antigens ; Clinical trials ; Coronaviruses ; COVID-19 ; Enzyme-linked immunosorbent assay ; Immune response (humoral) ; Immunity (humoral) ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Pandemics ; Severe acute respiratory syndrome coronavirus 2 ; Vaccines
Is Part Of: bioRxiv, 2021-07
Description: In response to the SARS-CoV-2 pandemic many vaccines have been developed and evaluated in human clinical trials. The humoral immune response magnitude, composition and efficacy of neutralizing SARS-CoV-2 are essential endpoints for these trials. Robust assays that are reproducibly precise, linear, and specific for SARS-CoV-2 antigens would be beneficial for the vaccine pipeline. In this work we describe the methodologies and clinical qualification of three SARS-CoV-2 endpoint assays. We developed and qualified Endpoint titer ELISAs for total IgG, IgG1, IgG3, IgG4, IgM and IgA to evaluate the magnitude of specific responses to the trimeric spike (S) antigen and total IgG specific to the spike receptor binding domain (RBD) of SARS-CoV-2. We also qualified a pseudovirus neutralization assay which evaluates functional antibody titers capable of inhibiting the entry and replication of a lentivirus containing the Spike antigen of SARS-CoV-2. To complete the suite of assays we qualified a plaque reduction neutralization test (PRNT) methodology using the 2019-nCoV/USA-WA1/2020 isolate of SARS-CoV-2 to assess neutralizing titers of antibodies in plasma from normal healthy donors and convalescent COVID-19 individuals. Competing Interest Statement The authors have declared no competing interest.
Publisher: Cold Spring Harbor: Cold Spring Harbor Laboratory Press
Language: English
Identifier: DOI: 10.1101/2021.07.02.450915
Source: Coronavirus Research Database

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Qualification of ELISA and neutralization methodologies to measure SARS-CoV-2 humoral immunity using human clinical samples

2021. This article is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (“the License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. ;DOI: 10.1101/2021.07.02.450915

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