EBioMedicine (May 2021)
A comprehensive antigen production and characterisation study for easy-to-implement, specific and quantitative SARS-CoV-2 serotests
- Miriam Klausberger,
- Mark Duerkop,
- Helmuth Haslacher,
- Gordana Wozniak-Knopp,
- Monika Cserjan-Puschmann,
- Thomas Perkmann,
- Nico Lingg,
- Patricia Pereira Aguilar,
- Elisabeth Laurent,
- Jelle De Vos,
- Manuela Hofner,
- Barbara Holzer,
- Maria Stadler,
- Gabriele Manhart,
- Klemens Vierlinger,
- Margot Egger,
- Lisa Milchram,
- Elisabeth Gludovacz,
- Nicolas Marx,
- Christoph Köppl,
- Christopher Tauer,
- Jürgen Beck,
- Daniel Maresch,
- Clemens Grünwald-Gruber,
- Florian Strobl,
- Peter Satzer,
- Gerhard Stadlmayr,
- Ulrike Vavra,
- Jasmin Huber,
- Markus Wahrmann,
- Farsad Eskandary,
- Marie-Kathrin Breyer,
- Daniela Sieghart,
- Peter Quehenberger,
- Gerda Leitner,
- Robert Strassl,
- Alexander E. Egger,
- Christian Irsara,
- Andrea Griesmacher,
- Gregor Hoermann,
- Günter Weiss,
- Rosa Bellmann-Weiler,
- Judith Loeffler-Ragg,
- Nicole Borth,
- Richard Strasser,
- Alois Jungbauer,
- Rainer Hahn,
- Jürgen Mairhofer,
- Boris Hartmann,
- Nikolaus B. Binder,
- Gerald Striedner,
- Lukas Mach,
- Andreas Weinhäusel,
- Benjamin Dieplinger,
- Florian Grebien,
- Wilhelm Gerner,
- Christoph J. Binder,
- Reingard Grabherr
Affiliations
- Miriam Klausberger
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Mark Duerkop
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; Novasign GmbH Vienna, Austria
- Helmuth Haslacher
- Department of Laboratory Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria
- Gordana Wozniak-Knopp
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; CD Laboratory for innovative Immunotherapeutics, Vienna, Austria
- Monika Cserjan-Puschmann
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Thomas Perkmann
- Department of Laboratory Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria
- Nico Lingg
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Patricia Pereira Aguilar
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Elisabeth Laurent
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; BOKU Core Facility Biomolecular & Cellular Analysis, University of Natural Resources and Life Sciences (BOKU),Vienna, Austria
- Jelle De Vos
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; Department of Chemical Engineering, Vrije Universiteit Brussel (VUB), Brussels, Belgium
- Manuela Hofner
- Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria
- Barbara Holzer
- Austrian Agency for Health and Food Safety (AGES), Department for Animal Health, Moedling, Austria
- Maria Stadler
- Institute of Immunology, University of Veterinary Medicine, Vienna, Austria
- Gabriele Manhart
- Institute for Medical Biochemistry, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria
- Klemens Vierlinger
- Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria
- Margot Egger
- Department of Laboratory Medicine, Konventhospital Barmherzige Brueder Linz and Ordensklinikum Linz Barmherzige Schwestern, Linz, Austria
- Lisa Milchram
- Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria
- Elisabeth Gludovacz
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Nicolas Marx
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Christoph Köppl
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Christopher Tauer
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Jürgen Beck
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Daniel Maresch
- BOKU Core Facility Mass Spectrometry, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria
- Clemens Grünwald-Gruber
- BOKU Core Facility Mass Spectrometry, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria; Department of Chemistry, University of Natural Resources and Life Sciences (BOKU) Vienna, Austria
- Florian Strobl
- enGenes Biotech GmbH, Vienna, Austria
- Peter Satzer
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Gerhard Stadlmayr
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; CD Laboratory for innovative Immunotherapeutics, Vienna, Austria
- Ulrike Vavra
- Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences (BOKU) Vienna, Austria
- Jasmin Huber
- Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria
- Markus Wahrmann
- Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Austria
- Farsad Eskandary
- Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Austria
- Marie-Kathrin Breyer
- Department of Respiratory and Critical Care Medicine and Ludwig Boltzmann Institute for Lung Health, Otto Wagner Hospital, Vienna, Austria
- Daniela Sieghart
- Division of Rheumatology, Department of Medicine III, Medical University of Vienna, Austria
- Peter Quehenberger
- Department of Laboratory Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria
- Gerda Leitner
- Department of Blood Group Serology and Transfusion Medicine, Medical University of Vienna, Austria
- Robert Strassl
- Department of Laboratory Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria
- Alexander E. Egger
- Central Institute for Medical and Chemical Laboratory Diagnosis, Innsbruck University Hospital, Innsbruck, Austria
- Christian Irsara
- Central Institute for Medical and Chemical Laboratory Diagnosis, Innsbruck University Hospital, Innsbruck, Austria
- Andrea Griesmacher
- Central Institute for Medical and Chemical Laboratory Diagnosis, Innsbruck University Hospital, Innsbruck, Austria
- Gregor Hoermann
- Central Institute for Medical and Chemical Laboratory Diagnosis, Innsbruck University Hospital, Innsbruck, Austria; MLL Munich Leukemia Laboratory, Munich, Germany
- Günter Weiss
- Department of Internal Medicine II, Innsbruck Medical University, Innsbruck, Austria
- Rosa Bellmann-Weiler
- Department of Internal Medicine II, Innsbruck Medical University, Innsbruck, Austria
- Judith Loeffler-Ragg
- Department of Internal Medicine II, Innsbruck Medical University, Innsbruck, Austria
- Nicole Borth
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria
- Richard Strasser
- Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences (BOKU) Vienna, Austria
- Alois Jungbauer
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Rainer Hahn
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria
- Jürgen Mairhofer
- enGenes Biotech GmbH, Vienna, Austria
- Boris Hartmann
- Austrian Agency for Health and Food Safety (AGES), Department for Animal Health, Moedling, Austria
- Nikolaus B. Binder
- Technoclone Herstellung von Diagnostika und Arzneimitteln GmbH, Vienna, Austria
- Gerald Striedner
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; Novasign GmbH Vienna, Austria; ACIB-Austrian Centre of Industrial Biotechnology, Graz, Austria; enGenes Biotech GmbH, Vienna, Austria
- Lukas Mach
- Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences (BOKU) Vienna, Austria
- Andreas Weinhäusel
- Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria
- Benjamin Dieplinger
- Department of Laboratory Medicine, Konventhospital Barmherzige Brueder Linz and Ordensklinikum Linz Barmherzige Schwestern, Linz, Austria
- Florian Grebien
- Institute for Medical Biochemistry, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria; Corresponding authors.
- Wilhelm Gerner
- Institute of Immunology, University of Veterinary Medicine, Vienna, Austria; Christian Doppler Laboratory for an Optimized Prediction of Vaccination Success in Pigs, University of Veterinary Medicine, Vienna, Austria; Present address: The Pirbright Institute, Pirbright, United Kingdom
- Christoph J. Binder
- Department of Laboratory Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria; Corresponding authors.
- Reingard Grabherr
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; Corresponding authors.
- Journal volume & issue
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Vol. 67
p. 103348
Abstract
Background: Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection or vaccination. While first-generation antibody tests have primarily provided qualitative results, accurate seroprevalence studies and tracking of antibody levels over time require highly specific, sensitive and quantitative test setups. Methods: We have developed two quantitative, easy-to-implement SARS-CoV-2 antibody tests, based on the spike receptor binding domain and the nucleocapsid protein. Comprehensive evaluation of antigens from several biotechnological platforms enabled the identification of superior antigen designs for reliable serodiagnostic. Cut-off modelling based on unprecedented large and heterogeneous multicentric validation cohorts allowed us to define optimal thresholds for the tests’ broad applications in different aspects of clinical use, such as seroprevalence studies and convalescent plasma donor qualification. Findings: Both developed serotests individually performed similarly-well as fully-automated CE-marked test systems. Our described sensitivity-improved orthogonal test approach assures highest specificity (99.8%); thereby enabling robust serodiagnosis in low-prevalence settings with simple test formats. 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 and disclosed antibody level thresholds that correlate well with robust neutralization of authentic SARS-CoV-2 virus. Interpretation: We demonstrate that antigen source and purity strongly impact serotest performance. Comprehensive biotechnology-assisted selection of antigens and in-depth characterisation of the assays allowed us to overcome limitations of simple ELISA-based antibody test formats based on chromometric reporters, to yield comparable assay performance as fully-automated platforms. Funding: WWTF, Project No. COV20–016; BOKU, LBI/LBG
