A comprehensive antigen production and characterization study for easy-to-implement, highly specific and quantitative SARS-CoV-2 antibody assays
Author:
Klausberger MiriamORCID, Dürkop MarkORCID, Haslacher HelmuthORCID, Wozniak-Knopp GordanaORCID, Cserjan-Puschmann MonikaORCID, Perkmann ThomasORCID, Lingg NicoORCID, Aguilar Patricia PereiraORCID, Laurent ElisabethORCID, De Vos JelleORCID, Hofer Manuela, Holzer Barbara, Stadler Maria, Manhart Gabriele, Vierlinger Klemens, Egger Margot, Milchram Lisa, Gludovacz Elisabeth, Marx NicolasORCID, Köppl Christoph, Tauer Christopher, Beck Jürgen, Maresch Daniel, Grünwald-Gruber Clemens, Strobl Florian, Satzer PeterORCID, Stadlmayr GerhardORCID, Vavra Ulrike, Huber Jasmin, Wahrmann MarkusORCID, Eskandary FarsadORCID, Breyer Marie-Kathrin, Sieghart DanielaORCID, Quehenberger PeterORCID, Leitner Gerda, Strassl RobertORCID, Egger Alexander E., Irsara ChristianORCID, Griesmacher Andrea, Hoermann GregorORCID, Weiss GünterORCID, Bellmann-Weiler Rosa, Loeffler-Ragg JudithORCID, Borth NicoleORCID, Strasser RichardORCID, Jungbauer AloisORCID, Hahn RainerORCID, Mairhofer JürgenORCID, Hartmann BorisORCID, Binder Nikolaus B., Striedner GeraldORCID, Mach LukasORCID, Weinhäusl AndreasORCID, Dieplinger Benjamin, Grebien FlorianORCID, Gerner WilhelmORCID, Binder Christoph J.ORCID, Grabherr ReingardORCID
Abstract
AbstractAntibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection. While first-generation antibody tests have primarily provided qualitative results with low specificity, accurate seroprevalence studies and tracking of antibody levels over time require highly specific, sensitive and quantitative test setups. Here, we describe two quantitative ELISA antibody tests based on the SARS-CoV-2 spike receptor-binding domain and the nucleocapsid protein. Comparative expression in bacterial, insect, mammalian and plant-based platforms enabled the identification of new antigen designs with superior quality and high suitability as diagnostic reagents. Both tests scored excellently in clinical validations with multi-centric specificity and sensitivity cohorts and showed unprecedented correlation with SARS-CoV-2 neutralization titers. Orthogonal testing increased assay specificity to 99.8%, thereby enabling robust serodiagnosis in low-prevalence settings. The inclusion of a calibrator permits accurate quantitative monitoring of antibody concentrations in samples collected at different time points during the acute and convalescent phase of COVID-19.
Publisher
Cold Spring Harbor Laboratory
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