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Qualification of ELISA and neutralization methodologies to measure SARS-CoV-2 humoral immunity using human clinical samples.
Larsen, Sasha E; Berube, Bryan J; Pecor, Tiffany; Cross, Evan; Brown, Bryan P; Williams, Brittany D; Johnson, Emma; Qu, Pingping; Carter, Lauren; Wrenn, Samuel; Kepl, Elizabeth; Sydeman, Claire; King, Neil P; Baldwin, Susan L; Coler, Rhea N.
  • Larsen SE; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Berube BJ; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America; HDT BioCorp., Seattle, WA, United States of America.
  • Pecor T; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Cross E; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Brown BP; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Williams BD; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America; Department of Global Health, University of Washington, Seattle, WA, United States of America.
  • Johnson E; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Qu P; Seattle Children's Research Institute, Biostatistics Epidemiology and Analytics in Research, Seattle, WA, United States of America.
  • Carter L; Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA 98195, United States of America.
  • Wrenn S; Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA 98195, United States of America.
  • Kepl E; Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA 98195, United States of America.
  • Sydeman C; Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA 98195, United States of America.
  • King NP; Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA 98195, United States of America.
  • Baldwin SL; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America.
  • Coler RN; Seattle Children's Research Institute, Center for Global Infectious Disease Research, Seattle, WA, United States of America; Department of Global Health, University of Washington, Seattle, WA, United States of America; Department of Pediatrics, University of Washington School of Medicine, Seattle, W
J Immunol Methods ; 499: 113160, 2021 12.
Article in English | MEDLINE | ID: covidwho-1440196
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ABSTRACT
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Enzyme-Linked Immunosorbent Assay / Neutralization Tests / Immunity, Humoral / SARS-CoV-2 Type of study: Experimental Studies / Prognostic study Topics: Vaccines Limits: Humans Language: English Journal: J Immunol Methods Year: 2021 Document Type: Article Affiliation country: J.jim.2021.113160

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Enzyme-Linked Immunosorbent Assay / Neutralization Tests / Immunity, Humoral / SARS-CoV-2 Type of study: Experimental Studies / Prognostic study Topics: Vaccines Limits: Humans Language: English Journal: J Immunol Methods Year: 2021 Document Type: Article Affiliation country: J.jim.2021.113160