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Linear B-Cell Epitopes in the Spike and Nucleocapsid Proteins as Markers of SARS-CoV-2 Exposure and Disease Severity

38 Pages Posted: 12 Jun 2020

See all articles by Siti Naqiah Amrun

Siti Naqiah Amrun

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Cheryl Yi-Pin Lee

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Bernett Lee

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Siew-Wai Fong

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN); National University of Singapore (NUS) - Department of Biological Sciences

Barnaby E. Young

National Centre for Infectious Diseases, Singapore

Rhonda Sin-Ling Chee

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Nicholas Kim-Wah Yeo

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Anthony Torres-Ruesta

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Guillaume Carissimo

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Chek Meng Poh

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Zi Wei Chang

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Matthew Zirui Tay

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Yi-Hao Chan

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Mark I-Cheng Chen

National Centre for Infectious Diseases

Paul A. Tambyah

National Centre for Infectious Diseases

Shirin Kalimuddin

Singapore General Hospital - Department of Infectious Diseases; Duke NUS Medical School - Emerging Infectious Disease Program

Surinder Pada

Ng Teng Fong General Hospital - Division of Infectious Diseases

Seow Yen Tan

Changi General Hospital

Louisa Jin Sun

Alexandra Hospital

Yee-Sin Leo

National University of Singapore (NUS) - Department of Microbiology and Immunology

David Chien Lye

National Centre for Infectious Diseases

Laurent Renia

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

Lisa FP Ng

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

More...

Abstract

Background: Given the unceasing worldwide surge in COVID-19 cases, there is an imperative need to develop highly specific and sensitive serology assays to define exposure to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

Methods: Pooled plasma samples from PCR positive COVID-19 patients were used to identify linear B-cell epitopes from a SARS-CoV-2 peptide library of spike (S), envelope (E), membrane (M), and nucleocapsid (N) structural proteins by peptide-based ELISA. Hit epitopes were further validated with 79 COVID-19 patients with different disease severity status, 12 seasonal human CoV, 20 recovered SARS patients and 22 healthy donors.

Findings: Four immunodominant epitopes, S14P5, S20P2, S21P2 and N4P5, were identified on the S and N viral proteins. IgG responses to all identified epitopes displayed a strong detection profile, with N4P5 achieving the highest level of specificity (100%) and sensitivity (>96%) against SARS-CoV-2. Furthermore, the magnitude of IgG responses to S14P5, S21P2 and N4P5 were strongly associated with disease severity.

Interpretation: IgG responses to the peptide epitopes can serve as useful indicators for the degree of immunopathology in COVID-19 patients, and function as higly specific and sensitive sero-immunosurveillance tools for recent or past SARS-CoV-2 infections. The flexibility of these epitopes to be used alone or in combination will allow for the development of improved point-of-care-tests (POCTs).

Funding Statement: Biomedical Research Council (BMRC), the A*ccelerate GAP-funded project (ACCL/19-GAP064-R20H-H) from Agency of Science, Technology and Research (A*STAR), and National Medical Research Council (NMRC) COVID-19 Research fund (COVID19RF-001). ATR is supported by the Singapore International Graduate Award (SINGA), A*STAR.

Declaration of Interests: The authors declare no conflict of interest.

Ethics Approval Statement: The study design and protocols for COVID-19, recovered SARS and seasonal hCoV patient cohorts were evaluated by National Healthcare Group (NHG) Domain Specific Review Board (DSRB) and approved under study numbers 2012/00917, 2020/00091 and 2020/00076, respectively. Healthy donor samples were collected under study numbers 2017/2806 and NUS IRB 04-140. Residual serum sample from National Healthy Survey (NHS) collected in 2010 under study number 006/2010 was used as a positive control in peptide-based ELISA (13). Written informed consent was obtained from participants in accordance with the Declaration of Helsinki for Human Research.

Keywords: epitopes; SARS-CoV-2; COVID-19; patients; biomarkers

Suggested Citation

Naqiah Amrun, Siti and Lee, Cheryl Yi-Pin and Lee, Bernett and Fong, Siew-Wai and Young, Barnaby E. and Sin-Ling Chee, Rhonda and Kim-Wah Yeo, Nicholas and Torres-Ruesta, Anthony and Carissimo, Guillaume and Meng Poh, Chek and Wei Chang, Zi and Zirui Tay, Matthew and Chan, Yi-Hao and Chen, Mark I-Cheng and Tambyah, Paul A. and Kalimuddin, Shirin and Pada, Surinder and Tan, Seow Yen and Jin Sun, Louisa and Leo, Yee-Sin and Lye, David Chien and Renia, Laurent and Ng, Lisa FP, Linear B-Cell Epitopes in the Spike and Nucleocapsid Proteins as Markers of SARS-CoV-2 Exposure and Disease Severity (5/16/2020). Available at SSRN: https://ssrn.com/abstract=3605114 or http://dx.doi.org/10.2139/ssrn.3605114

Siti Naqiah Amrun

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Cheryl Yi-Pin Lee

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Bernett Lee

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN) ( email )

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Siew-Wai Fong

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

National University of Singapore (NUS) - Department of Biological Sciences

14 Science Drive 4
Singapore, 117543
China

Barnaby E. Young

National Centre for Infectious Diseases, Singapore ( email )

Singapore

Rhonda Sin-Ling Chee

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Nicholas Kim-Wah Yeo

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Anthony Torres-Ruesta

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Guillaume Carissimo

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Chek Meng Poh

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Zi Wei Chang

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Matthew Zirui Tay

Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR)

Yi-Hao Chan

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Mark I-Cheng Chen

National Centre for Infectious Diseases

Paul A. Tambyah

National Centre for Infectious Diseases

Shirin Kalimuddin

Singapore General Hospital - Department of Infectious Diseases

Singapore

Duke NUS Medical School - Emerging Infectious Disease Program

Singapore

Surinder Pada

Ng Teng Fong General Hospital - Division of Infectious Diseases

Singapore

Seow Yen Tan

Changi General Hospital

Singapore

Louisa Jin Sun

Alexandra Hospital

Yee-Sin Leo

National University of Singapore (NUS) - Department of Microbiology and Immunology ( email )

5 Science Drive 2 Blk MD4, Level 3
117545
Singapore

David Chien Lye

National Centre for Infectious Diseases

Singapore

Laurent Renia

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

Lisa FP Ng (Contact Author)

Agency for Science, Technology and Research (A*STAR) - Singapore Immunology Network (SIgN)

1 Fusionopolis Way
#16-16 Connexis
Singapore, 138632
Singapore

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