Journal article
Novel ELISA Protocol Links Pre-Existing SARS-CoV-2 Reactive Antibodies With Endemic Coronavirus Immunity and Age and Reveals Improved Serologic Identification of Acute COVID-19 via Multi-Parameter Detection
Frontiers in immunology, Vol.12, 614676
04/09/2021
DOI: 10.3389/fimmu.2021.614676
PMCID: PMC8062931
PMID: 33897682
Abstract
The COVID-19 pandemic has drastically impacted work, economy, and way of life. Sensitive measurement of SARS-CoV-2 specific antibodies would provide new insight into pre-existing immunity, virus transmission dynamics, and the nuances of SARS-CoV-2 pathogenesis. To date, existing SARS-CoV-2 serology tests have limited utility due to insufficient reliable detection of antibody levels lower than what is typically present after several days of symptoms. To measure lower quantities of SARS-CoV-2 IgM, IgG, and IgA with higher resolution than existing assays, we developed a new ELISA protocol with a distinct plate washing procedure and timed plate development
use of a standard curve. Very low optical densities from samples added to buffer coated wells at as low as a 1:5 dilution are reported using this 'BU ELISA' method. Use of this method revealed circulating SARS-CoV-2 receptor binding domain (RBD) and nucleocapsid protein (N) reactive antibodies (IgG, IgM, and/or IgA) in 44 and 100 percent of pre-pandemic subjects, respectively, and the magnitude of these antibodies tracked with antibody levels of analogous viral proteins from endemic coronavirus (eCoV) strains. The disease status (HIV, SLE) of unexposed subjects was not linked with SARS-CoV-2 reactive antibody levels; however, quantities were significantly lower in subjects over 70 years of age compared with younger counterparts. Also, we measured SARS-CoV-2 RBD- and N- specific IgM, IgG, and IgA antibodies from 29 SARS-CoV-2 infected individuals at varying disease states, including 10 acute COVID-19 hospitalized subjects with negative serology results by the EUA approved Abbott IgG chemiluminescent microparticle immunoassay. Measurements of SARS-CoV-2 RBD- and N- specific IgM, IgG, IgA levels measured by the BU ELISA revealed higher signal from 9 of the 10 Abbott test negative COVID-19 subjects than all pre-pandemic samples for at least one antibody specificity/isotype, implicating improved serologic identification of SARS-CoV-2 infection
multi-parameter, high sensitive antibody detection. We propose that this improved ELISA protocol, which is straightforward to perform, low cost, and uses readily available commercial reagents, is a useful tool to elucidate new information about SARS-CoV-2 infection and immunity and has promising implications for improved detection of all analytes measurable by this platform.
Details
- Title: Subtitle
- Novel ELISA Protocol Links Pre-Existing SARS-CoV-2 Reactive Antibodies With Endemic Coronavirus Immunity and Age and Reveals Improved Serologic Identification of Acute COVID-19 via Multi-Parameter Detection
- Creators
- Rachel R Yuen - Boston University School of MedicineDylan Steiner - Boston University School of MedicineRiley M F Pihl - Boston University School of MedicineElizabeth Chavez - Boston UniversityAlex Olson - Boston Medical CenterErika L Smith - Boston University School of MedicineLillia A Baird - Boston University School of MedicineFiliz Korkmaz - Boston UniversityPatricia Urick - Boston University School of MedicineManish Sagar - Boston UniversityJacob L Berrigan - Boston University School of MedicineSuryaram Gummuluru - Boston University School of MedicineRonald B Corley - Boston UniversityKaren Quillen - Boston UniversityAnna C Belkina - Boston UniversityGustavo Mostoslavsky - Boston UniversityIan R Rifkin - Boston UniversityYachana Kataria - Boston UniversityAmedeo J Cappione III - MilliporeSigma, Burlington, MA, United StatesWenda Gao - Antagen PharmaceuticalsNina H Lin - Boston Medical CenterNahid Bhadelia - Boston University School of MedicineJennifer E Snyder-Cappione - Boston University School of Medicine
- Resource Type
- Journal article
- Publication Details
- Frontiers in immunology, Vol.12, 614676
- DOI
- 10.3389/fimmu.2021.614676
- PMID
- 33897682
- PMCID
- PMC8062931
- ISSN
- 1664-3224
- eISSN
- 1664-3224
- Grant note
- UC7 AI095321 / NIAID NIH HHS
- Language
- English
- Date published
- 04/09/2021
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984696653002771
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