SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup

Daniel Stadlbauer; Fatima Amanat; Veronika Chromikova; Kaijun Jiang; Shirin Strohmeier; Guha Asthagiri Arunkumar; Jessica Tan; Disha Bhavsar; Christina Capuano; Ericka Kirkpatrick; Philip Meade; Ruhi Nichalle Brito; Catherine Teo; Meagan McMahon; Viviana Simon; Florian Krammer

doi:10.1002/cpmc.100

SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup

Daniel Stadlbauer, Fatima Amanat, Veronika Chromikova, Kaijun Jiang, Shirin Strohmeier, Guha Asthagiri Arunkumar, Jessica Tan, Disha Bhavsar, Christina Capuano, Ericka Kirkpatrick, Philip Meade, Ruhi Nichalle Brito, Catherine Teo, Meagan McMahon, Viviana Simon, Florian Krammer

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Abstract

In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories.

Original language	English
Article number	e100
Journal	Current Protocols in Microbiology
Volume	57
Issue number	1
DOIs	https://doi.org/10.1002/cpmc.100
State	Published - 1 Jun 2020

Keywords

COVID-19
COVID19
ELISA
SARS-CoV-2
protein expression
serological assay

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10.1002/cpmc.100

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Stadlbauer, D., Amanat, F., Chromikova, V., Jiang, K., Strohmeier, S., Arunkumar, G. A., Tan, J., Bhavsar, D., Capuano, C., Kirkpatrick, E., Meade, P., Brito, R. N., Teo, C., McMahon, M., Simon, V., & Krammer, F. (2020). SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup. Current Protocols in Microbiology, 57(1), Article e100. https://doi.org/10.1002/cpmc.100

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title = "SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup",

abstract = "In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories.",

keywords = "COVID-19, COVID19, ELISA, SARS-CoV-2, protein expression, serological assay",

author = "Daniel Stadlbauer and Fatima Amanat and Veronika Chromikova and Kaijun Jiang and Shirin Strohmeier and Arunkumar, {Guha Asthagiri} and Jessica Tan and Disha Bhavsar and Christina Capuano and Ericka Kirkpatrick and Philip Meade and Brito, {Ruhi Nichalle} and Catherine Teo and Meagan McMahon and Viviana Simon and Florian Krammer",

note = "Funding Information: We thank Dr. Raffael Nachbagauer (Icahn School for Medicine at Mount Sinai) and Dr. Aubree Gordon (University of Michigan) for critical reading and constructive comments. Development of this protocol was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C. Funding Information: We thank Dr. Raffael Nachbagauer (Icahn School for Medicine at Mount Sinai) and Dr. Aubree Gordon (University of Michigan) for critical reading and constructive comments. Development of this protocol was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C. Philanthropic donations in support of our work are much appreciated, since the reagents are shared free of charge with the scientific community. Please contact Vanesa Saric (vanesa.saric@mountsinai.org) for further information. Publisher Copyright: {\textcopyright} 2020 The Authors. Current Protocols in Microbiology published by Wiley Periodicals LLC",

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Stadlbauer, D, Amanat, F, Chromikova, V, Jiang, K, Strohmeier, S, Arunkumar, GA, Tan, J, Bhavsar, D, Capuano, C, Kirkpatrick, E, Meade, P, Brito, RN, Teo, C, McMahon, M, Simon, V & Krammer, F 2020, 'SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup', Current Protocols in Microbiology, vol. 57, no. 1, e100. https://doi.org/10.1002/cpmc.100

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AU - Amanat, Fatima

AU - Chromikova, Veronika

AU - Jiang, Kaijun

AU - Strohmeier, Shirin

AU - Arunkumar, Guha Asthagiri

AU - Tan, Jessica

AU - Bhavsar, Disha

AU - Capuano, Christina

AU - Kirkpatrick, Ericka

AU - Meade, Philip

AU - Brito, Ruhi Nichalle

AU - Teo, Catherine

AU - McMahon, Meagan

AU - Simon, Viviana

AU - Krammer, Florian

N1 - Funding Information: We thank Dr. Raffael Nachbagauer (Icahn School for Medicine at Mount Sinai) and Dr. Aubree Gordon (University of Michigan) for critical reading and constructive comments. Development of this protocol was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C. Funding Information: We thank Dr. Raffael Nachbagauer (Icahn School for Medicine at Mount Sinai) and Dr. Aubree Gordon (University of Michigan) for critical reading and constructive comments. Development of this protocol was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C. Philanthropic donations in support of our work are much appreciated, since the reagents are shared free of charge with the scientific community. Please contact Vanesa Saric (vanesa.saric@mountsinai.org) for further information. Publisher Copyright: © 2020 The Authors. Current Protocols in Microbiology published by Wiley Periodicals LLC

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AB - In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories.

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DO - 10.1002/cpmc.100

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SN - 1934-8525

VL - 57

JO - Current Protocols in Microbiology

JF - Current Protocols in Microbiology

IS - 1

M1 - e100

ER -

SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup

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