Abstract

Serological assays have been widely used to detect anti-SARS-CoV-2 antibodies, which are generated from previous exposure to the virus or after vaccination. The presence of anti-SARS-CoV-2 Nucleocapsid antibodies was recently reported in patients´ urine using an in-house urine-based ELISA-platform, allowing a non-invasive way to collect clinical samples and assess immune conversion. In the current study, we evaluated and validated another in-house urine-based ELISA for the detection of anti-SARS-CoV-2 Spike antibodies. Three partial recombinant SARS-CoV-2 Spike proteins comprising the Receptor Binding Domain, expressed in eukaryotic or prokaryotic systems, were tested in an ELISA platform against a panel of over 140 urine and paired serum samples collected from 106 patients confirmed positive for SARS-CoV-2 by qRT-PCR. The key findings from our study were that anti-SARS-CoV-2 Spike antibodies could be detected in urine samples and that the prokaryotic expression of the rSARS-CoV-2 Spike protein was not a barrier to obtain relatively high serology efficiency for the urine-based assay. Thus, use of a urine-based ELISA assay with partial rSARS-CoV-2 Spike proteins, expressed in a prokaryotic system, could be considered as a convenient tool for screening for the presence of anti-SARS-CoV-2 Spike antibodies, and overcome the difficulties arising from sample collection and the need for recombinant proteins produced with eukaryotic expression systems.

Details

Title
A urine-based ELISA with recombinant non-glycosylated SARS-CoV-2 spike protein for detecting anti-SARS-CoV-2 spike antibodies
Author
Ramos, Fernanda F. 1 ; Bagno, Flávia F. 2 ; Vassallo, Paula F. 3 ; Oliveira-da-Silva, João A. 1 ; Reis, Thiago A. R. 1 ; Bandeira, Raquel S. 1 ; Machado, Amanda S. 1 ; Lage, Daniela P. 4 ; Martins, Vivian T. 1 ; Fernandes, Ana P. 5 ; Christodoulides, Myron 6 ; Ravetti, Cecilia G. 7 ; Nobre, Vandack 8 ; da Fonseca, Flávio G. 9   VIAFID ORCID Logo  ; Coelho, Eduardo A. F. 4 ; Ludolf, Fernanda 1   VIAFID ORCID Logo 

1  Universidade Federal de Minas Gerais, Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
2  Universidade Federal de Minas Gerais, Centro de Tecnologia de Vacinas (CT Vacinas) / BH-Tec, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
3  Universidade Federal de Minas Gerais, Hospital das Clínicas, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
4  Universidade Federal de Minas Gerais, Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888); Universidade Federal de Minas Gerais, Departamento de Patologia Clínica, COLTEC, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
5  Universidade Federal de Minas Gerais, Centro de Tecnologia de Vacinas (CT Vacinas) / BH-Tec, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888); Universidade Federal de Minas Gerais, Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
6  University of Southampton Faculty of Medicine, Neisseria Research Group, School of Clinical and Experimental Sciences, Southampton, England (GRID:grid.5491.9) (ISNI:0000 0004 1936 9297) 
7  Universidade Federal de Minas Gerais, Departamento de Clínica Médica, Faculdade de Medicina, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
8  Universidade Federal de Minas Gerais, Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888); Universidade Federal de Minas Gerais, Departamento de Clínica Médica, Faculdade de Medicina, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
9  Universidade Federal de Minas Gerais, Centro de Tecnologia de Vacinas (CT Vacinas) / BH-Tec, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888); Universidade Federal de Minas Gerais, Laboratório de Virologia Molecular e Aplicada, Departamento de Microbiologia, Instituto de Ciências Biológicas, Belo Horizonte, Brazil (GRID:grid.8430.f) (ISNI:0000 0001 2181 4888) 
Pages
4345
Publication year
2023
Publication date
2023
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-023-31382-5
ProQuest document ID
2787393533
Copyright
© The Author(s) 2023. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.