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Sensitive Detection of Total Anti-Spike Antibodies and Isotype Switching in Asymptomatic and Symptomatic COVID-19 Patients

44 Pages Posted: 23 Oct 2020 Publication Status: Published

See all articles by Yun Shan Goh

Yun Shan Goh

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre

Jean-Marc Chavatte

National Centre for Infectious Diseases

Alicia Lim Jieling

National Centre for Infectious Diseases

Bernett Lee

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

Siti Naqiah Amrun

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre

Cheryl Yi-Pin Lee

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

Rhonda Sin-Ling Chee

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre

Bei Wang

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

Chia Yin Lee

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

Eve Ngoh

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

Cheng-I Wang

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

Barnaby E. Young

National Centre for Infectious Diseases, Singapore

Paul A. Tambyah

National Centre for Infectious Diseases

Shirin Kalimuddin

National University of Singapore (NUS) - 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 - Department of Infectious Diseases

Louisa Sun

Alexandra Hospital

Mark I-Cheng Chen

National Centre for Infectious Diseases

Yee-Sin Leo

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

David Chien Lye

Nanyang Technological University (NTU) - Lee Kong Chian School of Medicine; National University of Singapore (NUS) - Department of Microbiology and Immunology

Lisa F.P. Ng

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

Raymond Tzer Pin Lin

National Centre for Infectious Diseases

Laurent Renia

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre

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Abstract

Early detection of infections is crucial to limit the spread of coronavirus 2019 disease (COVID-19). Here, we developed a flow cytometry-based assay to detect SARS-CoV-2 Spike protein (S protein) antibodies in COVID-19 patients. The assay detected specific IgM and IgG in COVID-19 patients and also the acquisition of all IgG subclasses, with IgG1 being the most dominant. The antibody response was significantly higher at a later stage of the infection. Furthermore, asymptomatic COVID-19 patients also developed specific IgM and IgG, with IgG1 as the most dominant subclass. Although the antibody levels were lower in asymptomatic infections, the assay was highly sensitive and detected 97% of asymptomatic infections. These findings demonstrated that the assay could be used for serological analysis of symptomatic patients, and also as a sensitive tool to detect asymptomatic infections, which may go undetected.

Funding: 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, COVID-19RF-007, COVID-19RF-60).

Conflict of Interest: The authors declare no competing interests.

Ethical Approval: The study design and protocols for COVID-19, recovered SARS and seasonal human CoV patient cohorts were approved by National Healthcare Group (NHG) Domain Specific Review Board (DSRB) and performed, following ethical guidelines in the approved studies 2012/00917, 2020/00091 and 2020/00076 respectively. Healthy donor samples were collected in accordance with approved studies 2017/2806 and NUS IRB 04-140. Written informed consent was obtained from participants in accordance with the Declaration of Helsinki for Human Research.

Keywords: SARS-CoV-2, COVID-19, serological, S protein, antibodies, IgG subclasses, symptomatic, asymptomatic

Suggested Citation

Goh, Yun Shan and Chavatte, Jean-Marc and Jieling, Alicia Lim and Lee, Bernett and Amrun, Siti Naqiah and Lee, Cheryl Yi-Pin and Chee, Rhonda Sin-Ling and Wang, Bei and Lee, Chia Yin and Ngoh, Eve and Wang, Cheng-I and Young, Barnaby E. and Tambyah, Paul A. and Kalimuddin, Shirin and Pada, Surinder and Tan, Seow-Yen and Sun, Louisa and Chen, Mark I-Cheng and Leo, Yee-Sin and Lye, David Chien and Ng, Lisa F.P. and Lin, Raymond Tzer Pin and Renia, Laurent, Sensitive Detection of Total Anti-Spike Antibodies and Isotype Switching in Asymptomatic and Symptomatic COVID-19 Patients. Available at SSRN: https://ssrn.com/abstract=3713507 or http://dx.doi.org/10.2139/ssrn.3713507
This version of the paper has not been formally peer reviewed.

Yun Shan Goh

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre ( email )

Singapore

Jean-Marc Chavatte

National Centre for Infectious Diseases ( email )

Singapore

Alicia Lim Jieling

National Centre for Infectious Diseases

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

Siti Naqiah Amrun

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre ( email )

Singapore

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

Rhonda Sin-Ling Chee

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre ( email )

Singapore

Bei Wang

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

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

Chia Yin Lee

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

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

Eve Ngoh

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

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

Cheng-I Wang

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

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

Barnaby E. Young

National Centre for Infectious Diseases, Singapore ( email )

Singapore

Paul A. Tambyah

National Centre for Infectious Diseases

Singapore

Shirin Kalimuddin

National University of Singapore (NUS) - 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 - Department of Infectious Diseases ( email )

Louisa Sun

Alexandra Hospital

Singapore

Mark I-Cheng Chen

National Centre for Infectious Diseases

Singapore

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

Nanyang Technological University (NTU) - Lee Kong Chian School of Medicine ( email )

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

5 Science Drive 2 Blk MD4, Level 3
117545
Singapore

Lisa F.P. Ng

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

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

Raymond Tzer Pin Lin

National Centre for Infectious Diseases

Singapore

Laurent Renia (Contact Author)

Agency for Science, Technology and Research (A*STAR) - Infectious Diseases Horizontal Technology Centre ( email )

Singapore

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