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Systematic evaluation of SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0003-0880-5375
KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics.ORCID iD: 0000-0002-8593-9089
KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics.
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Number of Authors: 502021 (English)In: Clinical & Translational Immunology (CTI), E-ISSN 2050-0068, Vol. 10, no 7, article id e1312Article in journal (Refereed) Published
Abstract [en]

Objective. The COVID-19 pandemic poses an immense need for accurate, sensitive and high-throughput clinical tests, and serological assays are needed for both overarching epidemiological studies and evaluating vaccines. Here, we present the development and validation of a high-throughput multiplex bead-based serological assay. Methods. More than 100 representations of SARS-CoV-2 proteins were included for initial evaluation, including antigens produced in bacterial and mammalian hosts as well as synthetic peptides. The five best-performing antigens, three representing the spike glycoprotein and two representing the nucleocapsid protein, were further evaluated for detection of IgG antibodies in samples from 331 COVID-19 patients and convalescents, and in 2090 negative controls sampled before 2020. Results. Three antigens were finally selected, represented by a soluble trimeric form and the S1-domain of the spike glycoprotein as well as by the C-terminal domain of the nucleocapsid. The sensitivity for these three antigens individually was found to be 99.7%, 99.1% and 99.7%, and the specificity was found to be 98.1%, 98.7% and 95.7%. The best assay performance was although achieved when utilising two antigens in combination, enabling a sensitivity of up to 99.7% combined with a specificity of 100%. Requiring any two of the three antigens resulted in a sensitivity of 99.7% and a specificity of 99.4%. Conclusion. These observations demonstrate that a serological test based on a combination of several SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay.

Place, publisher, year, edition, pages
WILEY , 2021. Vol. 10, no 7, article id e1312
Keywords [en]
COVID-19, IgG, multiplex, SARS-CoV-2, serological assay
National Category
Infectious Medicine
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URN: urn:nbn:se:kth:diva-299708DOI: 10.1002/cti2.1312ISI: 000680165000008PubMedID: 34295471Scopus ID: 2-s2.0-85111256975OAI: oai:DiVA.org:kth-299708DiVA, id: diva2:1585159
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QC 20210816

Available from: 2021-08-16 Created: 2021-08-16 Last updated: 2024-03-15Bibliographically approved

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Hober, SophiaHellström, CeciliaOlofsson, JennieAndersson, EniBergström, SofiaJernbom Falk, AugustBayati, ShaghayeghMravinacová, SáraSjöberg, RonaldYousef, JamilSkoglund, LovisaKanje, SaraBerling, AnnaSvensson, Anne-SophieJensen, GabriellaEnstedt, HenricAfshari, DelaramXu, Lan LanZwahlen, Martinvon Feilitzen, KalleLendel, ChristoferSivertsson, ÅsaTegel, HannaPin, ElisaMånberg, AnnaHedhammar, MyNilsson, Peter

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Hober, SophiaHellström, CeciliaOlofsson, JennieAndersson, EniBergström, SofiaJernbom Falk, AugustBayati, ShaghayeghMravinacová, SáraSjöberg, RonaldYousef, JamilSkoglund, LovisaKanje, SaraBerling, AnnaSvensson, Anne-SophieJensen, GabriellaEnstedt, HenricAfshari, DelaramXu, Lan LanZwahlen, Martinvon Feilitzen, KalleLendel, ChristoferSivertsson, ÅsaTegel, HannaPin, ElisaMånberg, AnnaHedhammar, MyNilsson, Peter
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Affinity ProteomicsScience for Life Laboratory, SciLifeLabProtein ScienceProtein TechnologySystems BiologyApplied Physical ChemistryCentre for Bioprocess Technology, CBioPT
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