B-Cell Epitopes-Based Chimeric Protein from SARS-CoV-2 N and S Proteins Is Recognized by Specific Antibodies in Serum and Urine Samples from Patients

Author:

Ramos Fernanda F.1ORCID,Pereira Isabela A. G.1,Cardoso Mariana M.1,Bandeira Raquel S.1,Lage Daniela P.1,Scussel Rahisa2ORCID,Anastacio Rafaela S.2,Freire Victor G.2,Melo Marina F. N.1,Oliveira-da-Silva Joao A.1,Martins Vivian T.1,Tavares Grasiele S. V.1,Vale Danniele L.1,Freitas Camila S.1ORCID,Chaves Ana Thereza1,Caporali Júlia F. M.13ORCID,Vassallo Paula F.3,Ravetti Cecilia G.3ORCID,Nobre Vandack13ORCID,Fonseca Flavio G.45ORCID,Christodoulides Myron6ORCID,Machado-de-Ávila Ricardo A.2ORCID,Coelho Eduardo A. F.17ORCID,Ludolf Fernanda18ORCID

Affiliation:

1. Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, Minas Gerais, Brazil

2. Programa de Pós-Graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma 88806-000, Santa Catarina, Brazil

3. Departamento de Clínica Médica, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, Minas Gerais, Brazil

4. Centro de Tecnologia de Vacinas (CT Vacinas), BH-Tec, UFMG, Belo Horizonte 31270-901, Minas Gerais, Brazil

5. Laboratório de Virologia Molecular e Aplicada, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Minas Gerais, Brazil

6. Neisseria Research Group, Molecular Microbiology, School of Clinical and Experimental Sciences, University of Southampton Faculty of Medicine, Southampton General Hospital, Southampton SO16 6YD, UK

7. Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Minas Gerais, Brazil

8. Programa de Pós-Graduação em Ciências da Saúde, Faculdade Ciências Médicas de Minas Gerais, Belo Horizonte 30130-110, Minas Gerais, Brazil

Abstract

The impact of the COVID-19 pandemic caused by the SARS-CoV-2 virus underscored the crucial role of laboratorial tests as a strategy to control the disease, mainly to indicate the presence of specific antibodies in human samples from infected patients. Therefore, suitable recombinant antigens are relevant for the development of reliable tests, and so far, single recombinant proteins have been used. In this context, B-cell epitopes-based chimeric proteins can be an alternative to obtain tests with high accuracy through easier and cheaper production. The present study used bioinformatics tools to select specific B-cell epitopes from the spike (S) and the nucleocapsid (N) proteins from the SARS-CoV-2 virus, aiming to produce a novel recombinant chimeric antigen (N4S11-SC2). Eleven S and four N-derived B-cell epitopes were predicted and used to construct the N4S11-SC2 protein, which was analyzed in a recombinant format against serum and urine samples, by means of an in house-ELISA. Specific antibodies were detected in the serum and urine samples of COVID-19 patients, which were previously confirmed by qRT-PCR. Results showed that N4S11-SC2 presented 83.7% sensitivity and 100% specificity when using sera samples, and 91.1% sensitivity and 100% specificity using urine samples. Comparable findings were achieved with paired urine samples when compared to N and S recombinant proteins expressed in prokaryotic systems. However, better results were reached for N4S11-SC2 in comparison to the S recombinant protein when using paired serum samples. Anti-N4S11-SC2 antibodies were not clearly identified in Janssen Ad26.COV2.S COVID-19-vaccinated subjects, using serum or paired urine samples. In conclusion, this study presents a new chimeric recombinant antigen expressed in a prokaryotic system that could be considered as an alternative diagnostic marker for the SARS-CoV-2 infection, with the potential benefits to be used on serum or urine from infected patients.

Funder

Secretaria de Educação Superior do Ministério da Educação

Fundação de Amparo à Pesquisa do Estado de Minas Gerais

Brazilian agencies Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Publisher

MDPI AG

Subject

Virology,Infectious Diseases

Reference34 articles.

1. FDA (2022, August 22). EUA Authorized Serology Test Performance. Food and Drug Administration, Available online: https://www.fda.gov/medical-devices/coronavirus-disease-2019-covid-19-emergency-use-authorizations-medical-devices/eua-authorized-serology-test-performance.

2. CDC (2023, August 16). COVID-19 Testing: What You Need to Know, Available online: https://www.cdc.gov/coronavirus/2019-ncov/symptoms-testing/testing.html.

3. Association of Self-reported COVID-19 Infection and SARS-CoV-2 Serology Test Results with Persistent Physical Symptoms among French Adults during the COVID-19 Pandemic;Matta;JAMA Intern. Med.,2022

4. How to interpret and use COVID-19 serology and immunology tests;Ong;Clin. Microbiol. Infect.,2021

5. FDA (2023, August 16). SARS-CoV-2 Viral Mutations: Impact on COVID-19 Tests. Food and Drug Administration, Available online: https://www.fda.gov/medical-devices/coronavirus-covid-19-and-medical-devices/sars-cov-2-viral-mutations-impact-covid-19-tests.

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