The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization

doi:10.1128/msphere.00193-22

. 2022 Aug 31;7(4):e0019322.

doi: 10.1128/msphere.00193-22. Epub 2022 Jun 15.

The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization

Amy B Karger¹, James D Brien², Jayne M Christen³, Santosh Dhakal⁴, Troy J Kemp³, Sabra L Klein⁴, Ligia A Pinto³, Lakshmanane Premkumar⁵, John D Roback⁶, Raquel A Binder⁷, Karl W Boehme⁸, Suresh Boppana^{9

10}, Carlos Cordon-Cardo¹¹, James M Crawford¹², John L Daiss¹³, Alan P Dupuis 2nd¹⁴, Ana M Espino¹⁵, Adolfo Firpo-Betancourt¹¹, Catherine Forconi⁷, J Craig Forrest⁸, Roxie C Girardin¹⁴, Douglas A Granger¹⁶, Steve W Granger¹⁶, Natalie S Haddad¹⁷, Christopher D Heaney¹⁸, Danielle T Hunt¹⁴, Joshua L Kennedy^{19

20

21}, Christopher L King²², Florian Krammer²³, Kate Kruczynski¹⁸, Joshua LaBaer²⁴, F Eun-Hyung Lee¹⁷, William T Lee^{14

25}, Shan-Lu Liu^{26

27

28

29}, Gerard Lozanski³⁰, Todd Lucas^{31

32}, Damodara Rao Mendu¹¹, Ann M Moormann⁷, Vel Murugan²⁴, Nkemakonam C Okoye¹², Petraleigh Pantoja³³, Anne F Payne¹⁴, Jin Park²⁴, Swetha Pinninti⁹, Amelia K Pinto², Nora Pisanic¹⁸, Ji Qiu²⁴, Carlos A Sariol^{33

34}, Viviana Simon²³, Lusheng Song²⁴, Tara L Steffen², E Taylor Stone², Linda M Styer^{14

25}, Mehul S Suthar^{35

36

37

38}, Stefani N Thomas¹, Bharat Thyagarajan¹, Ania Wajnberg³⁹, Jennifer L Yates^{14

25}, Kimia Sobhani⁴⁰

Affiliations

¹ Department of Laboratory Medicine and Pathology, University of Minnesotagrid.17635.36, Minneapolis, Minnesota, USA.
² Department of Molecular Microbiology & Immunology, Saint Louis Universitygrid.262962.b, St. Louis, Missouri, USA.
³ Frederick National Laboratory for Cancer Researchgrid.418021.e, Frederick, Maryland, USA.
⁴ W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA.
⁵ Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, USA.
⁶ Department of Pathology and Laboratory Medicine, grid.471395.dgrid.189967.8Emory University School of Medicinegrid.471395.d, Atlanta, Georgia, USA.
⁷ Department of Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁸ Department of Microbiology & Immunology, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
⁹ Department of Pediatrics, University of Alabama at Birminghamgrid.265892.2, Birmingham, Alabama, USA.
¹⁰ Department of Microbiology, University of Alabama at Birminghamgrid.265892.2, Birmingham, Alabama, USA.
¹¹ Department of Pathology, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
¹² Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York, USA.
¹³ MicroB-plex, Inc., Atlanta, Georgia, USA.
¹⁴ Wadsworth Centergrid.465543.5, New York State Department of Health, Albany, New York, USA.
¹⁵ Department of Microbiology and Medical Zoology, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
¹⁶ Salimetrics, LLC, Carlsbad, California, USA.
¹⁷ Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Department of Medicine, grid.471395.dgrid.189967.Emory University, Atlanta, Georgia, USA.
¹⁸ Department of Environmental Health and Engineering, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA.
¹⁹ Department of Pediatrics, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
²⁰ Department of Internal Medicine, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
²¹ Arkansas Children's Research Institute, Little Rock, Arkansas, USA.
²² Department of Pathology, Case Western Reserve School of Medicine, Cleveland, Ohio, USA.
²³ Department of Microbiology, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
²⁴ Virginia G. Piper Center for Personalized Diagnostics, Arizona State University Biodesign Institute, Tempe, Arizona, USA.
²⁵ Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, New York, USA.
²⁶ Center for Retrovirus Research, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁷ Department of Veterinary Biosciences, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁸ Department of Microbial Infection and Immunity, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁹ Viruses and Emerging Pathogens Program, Infectious Disease Institute, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
³⁰ Department of Pathology, The Ohio State Universitygrid.261331.4 Medical Center, Columbus, Ohio, USA.
³¹ Division of Public Health, College of Human Medicine, Michigan State University, East Lansing, Michigan, USA.
³² Department of Epidemiology, College of Human Medicine, Michigan State University, East Lansing, Michigan, USA.
³³ Unit of Comparative Medicine, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
³⁴ Department of Internal Medicine, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
³⁵ Center for Childhood Infections and Vaccines of Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicinegrid.471395.d, Atlanta, Georgia, USA.
³⁶ Department of Microbiology and Immunology, grid.471395.dgrid.189967.8Emory University, Atlanta, Georgia, USA.
³⁷ Emory Vaccine Center, grid.471395.dgrid.189967.8Emory University, Atlanta, Georgia, USA.
³⁸ Yerkes National Primate Research Center, Atlanta, Georgia, USA.
³⁹ Department of Medicine, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
⁴⁰ Department of Pathology and Laboratory Medicine, Cedars-Sinai Medical Centergrid.50956.3f, Los Angeles, California, USA.

PMID: 35703544
PMCID: PMC9429934
DOI: 10.1128/msphere.00193-22

The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization

Amy B Karger et al. mSphere. 2022.

. 2022 Aug 31;7(4):e0019322.

doi: 10.1128/msphere.00193-22. Epub 2022 Jun 15.

Authors

Affiliations

¹ Department of Laboratory Medicine and Pathology, University of Minnesotagrid.17635.36, Minneapolis, Minnesota, USA.
² Department of Molecular Microbiology & Immunology, Saint Louis Universitygrid.262962.b, St. Louis, Missouri, USA.
³ Frederick National Laboratory for Cancer Researchgrid.418021.e, Frederick, Maryland, USA.
⁴ W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA.
⁵ Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, USA.
⁶ Department of Pathology and Laboratory Medicine, grid.471395.dgrid.189967.8Emory University School of Medicinegrid.471395.d, Atlanta, Georgia, USA.
⁷ Department of Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁸ Department of Microbiology & Immunology, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
⁹ Department of Pediatrics, University of Alabama at Birminghamgrid.265892.2, Birmingham, Alabama, USA.
¹⁰ Department of Microbiology, University of Alabama at Birminghamgrid.265892.2, Birmingham, Alabama, USA.
¹¹ Department of Pathology, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
¹² Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York, USA.
¹³ MicroB-plex, Inc., Atlanta, Georgia, USA.
¹⁴ Wadsworth Centergrid.465543.5, New York State Department of Health, Albany, New York, USA.
¹⁵ Department of Microbiology and Medical Zoology, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
¹⁶ Salimetrics, LLC, Carlsbad, California, USA.
¹⁷ Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Department of Medicine, grid.471395.dgrid.189967.Emory University, Atlanta, Georgia, USA.
¹⁸ Department of Environmental Health and Engineering, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA.
¹⁹ Department of Pediatrics, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
²⁰ Department of Internal Medicine, University of Arkansas for Medical Sciencesgrid.241054.6, Little Rock, Arkansas, USA.
²¹ Arkansas Children's Research Institute, Little Rock, Arkansas, USA.
²² Department of Pathology, Case Western Reserve School of Medicine, Cleveland, Ohio, USA.
²³ Department of Microbiology, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
²⁴ Virginia G. Piper Center for Personalized Diagnostics, Arizona State University Biodesign Institute, Tempe, Arizona, USA.
²⁵ Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, New York, USA.
²⁶ Center for Retrovirus Research, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁷ Department of Veterinary Biosciences, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁸ Department of Microbial Infection and Immunity, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
²⁹ Viruses and Emerging Pathogens Program, Infectious Disease Institute, The Ohio State Universitygrid.261331.4, Columbus, Ohio, USA.
³⁰ Department of Pathology, The Ohio State Universitygrid.261331.4 Medical Center, Columbus, Ohio, USA.
³¹ Division of Public Health, College of Human Medicine, Michigan State University, East Lansing, Michigan, USA.
³² Department of Epidemiology, College of Human Medicine, Michigan State University, East Lansing, Michigan, USA.
³³ Unit of Comparative Medicine, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
³⁴ Department of Internal Medicine, University of Puerto Rico-Medical Sciences Campus, San Juan, Puerto Rico, USA.
³⁵ Center for Childhood Infections and Vaccines of Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicinegrid.471395.d, Atlanta, Georgia, USA.
³⁶ Department of Microbiology and Immunology, grid.471395.dgrid.189967.8Emory University, Atlanta, Georgia, USA.
³⁷ Emory Vaccine Center, grid.471395.dgrid.189967.8Emory University, Atlanta, Georgia, USA.
³⁸ Yerkes National Primate Research Center, Atlanta, Georgia, USA.
³⁹ Department of Medicine, Icahn School of Medicine at Mount Sinaigrid.59734.3c, New York, New York, USA.
⁴⁰ Department of Pathology and Laboratory Medicine, Cedars-Sinai Medical Centergrid.50956.3f, Los Angeles, California, USA.

PMID: 35703544
PMCID: PMC9429934
DOI: 10.1128/msphere.00193-22

Abstract

In October 2020, the National Cancer Institute (NCI) Serological Sciences Network (SeroNet) was established to study the immune response to COVID-19, and "to develop, validate, improve, and implement serological testing and associated technologies" (https://www.cancer.gov/research/key-initiatives/covid-19/coronavirus-research-initiatives/serological-sciences-network). SeroNet is comprised of 25 participating research institutions partnering with the Frederick National Laboratory for Cancer Research (FNLCR) and the SeroNet Coordinating Center. Since its inception, SeroNet has supported collaborative development and sharing of COVID-19 serological assay procedures and has set forth plans for assay harmonization. To facilitate collaboration and procedure sharing, a detailed survey was sent to collate comprehensive assay details and performance metrics on COVID-19 serological assays within SeroNet. In addition, FNLCR established a protocol to calibrate SeroNet serological assays to reference standards, such as the U.S. severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology standard reference material and first WHO international standard (IS) for anti-SARS-CoV-2 immunoglobulin (20/136), to facilitate harmonization of assay reporting units and cross-comparison of study data. SeroNet institutions reported development of a total of 27 enzyme-linked immunosorbent assay (ELISA) methods, 13 multiplex assays, and 9 neutralization assays and use of 12 different commercial serological methods. FNLCR developed a standardized protocol for SeroNet institutions to calibrate these diverse serological assays to reference standards. In conclusion, SeroNet institutions have established a diverse array of COVID-19 serological assays to study the immune response to SARS-CoV-2 and vaccines. Calibration of SeroNet serological assays to harmonize results reporting will facilitate future pooled data analyses and study cross-comparisons. IMPORTANCE SeroNet institutions have developed or implemented 61 diverse COVID-19 serological assays and are collaboratively working to harmonize these assays using reference materials to establish standardized reporting units. This will facilitate clinical interpretation of serology results and cross-comparison of research data.

Keywords: COVID-19; SeroNet; assay harmonization; serology.

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Conflict of interest statement

The authors declare a conflict of interest.

Figures

**FIG 1**
Example plate map for assay calibration setup. Numbers indicate suggested serial dilutions. Serial dilutions of primary and secondary calibrators (reference materials) are plated in triplicate, and the daily internal assay standard is plated in quadruplicate. C_STD, daily internal assay standard; STD-C1, -C2, and -C3, primary calibrator (primary reference material or standard); STD-T1, -T2, and T3, secondary calibrator (secondary reference material or standard); NEG, negative control sample; PC1, positive control sample 1; PC2, positive control sample 2.

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Update of

The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization.
Karger AB, Brien JD, Christen JM, Dhakal S, Kemp TJ, Klein SL, Pinto LA, Premkumar L, Roback JD, Binder RA, Boehme KW, Boppana S, Cordon-Cardo C, Crawford JM, Daiss JL, Dupuis AP 2nd, Espino AM, Firpo-Betancourt A, Forconi C, Forrest JC, Girardin RC, Granger DA, Granger SW, Haddad NS, Heaney CD, Hunt DT, Kennedy JL, King CL, Krammer F, Kruczynski K, LaBaer J, Lee FE, Lee WT, Liu SL, Lozanski G, Lucas T, Mendu DR, Moormann AM, Murugan V, Okoye NC, Pantoja P, Payne AF, Park J, Pinninti S, Pinto AK, Pisanic N, Qiu J, Sariol CA, Simon V, Song L, Steffen TL, Stone ET, Styer LM, Suthar MS, Thomas SN, Thyagarajan B, Wajnberg A, Yates JL, Sobhani K. Karger AB, et al. medRxiv [Preprint]. 2022 Mar 14:2022.02.27.22271399. doi: 10.1101/2022.02.27.22271399. medRxiv. 2022. Update in: mSphere. 2022 Aug 31;7(4):e0019322. doi: 10.1128/msphere.00193-22. PMID: 35262095 Free PMC article. Updated. Preprint.

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References

1. National Cancer Institute. 2022. NCI Serological Sciences Network for COVID-19 (SeroNet). https://www.cancer.gov/research/key-initiatives/covid-19/coronavirus-res.... Accessed 2 December 2021.
1. Food and Drug Administration (FDA). In vitro diagnostics EUAs—serology and other adaptive immune response tests for SARS-CoV-2. https://www.fda.gov/medical-devices/coronavirus-disease-2019-covid-19-em.... Accessed 2 December 2021.
1. Muecksch F, Wise H, Batchelor B, Squires M, Semple E, Richardson C, McGuire J, Clearly S, Furrie E, Greig N, Hay G, Templeton K, Lorenzi JCC, Hatziioannou T, Jenks S, Bieniasz PD. 2021. Longitudinal serological analysis and neutralizing antibody levels in coronavirus disease 2019 convalescent patients. J Infect Dis 223:389–398. doi:10.1093/infdis/jiaa659. - DOI - PMC - PubMed
1. Perez-Saez J, Zaballa M-E, Yerly S, Andrey DO, Meyer B, Eckerle I, Balavoine J-F, Chappuis F, Pittet D, Trono D, Kherad O, Vuilleumier N, Kaiser L, Guessous I, Stringhini S, Azman AS, Specchio-COVID19 Study Group . 2021. Persistence of anti-SARS-CoV-2 antibodies: immunoassay heterogeneity and implications for serosurveillance. Clin Microbiol Infect 27:1695.e7–e1695.e12. doi:10.1016/j.cmi.2021.06.040. - DOI - PMC - PubMed
1. Muecksch F, Wise H, Templeton K, Batchelor B, Squires M, McCance K, Jarvis L, Malloy K, Furrie E, Richardson C, MacGuire J, Godber I, Burns A, Mavin S, Zhang F, Schmidt F, Bieniasz P, Jenks S, Hatziioannou T. 2021. Longitudinal variation in SARS-CoV-2 antibody levels and emergence of viral variants: implications for the ability of serological assays to predict immunity. medRxiv doi:10.1101/2021.07.02.21259939. - DOI - PMC - PubMed

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[1] National Cancer Institute. 2022. NCI Serological Sciences Network for COVID-19 (SeroNet). https://www.cancer.gov/research/key-initiatives/covid-19/coronavirus-res.... Accessed 2 December 2021.

[2] National Cancer Institute. 2022. NCI Serological Sciences Network for COVID-19 (SeroNet). https://www.cancer.gov/research/key-initiatives/covid-19/coronavirus-res.... Accessed 2 December 2021.

[3] Food and Drug Administration (FDA). In vitro diagnostics EUAs—serology and other adaptive immune response tests for SARS-CoV-2. https://www.fda.gov/medical-devices/coronavirus-disease-2019-covid-19-em.... Accessed 2 December 2021.

[4] Food and Drug Administration (FDA). In vitro diagnostics EUAs—serology and other adaptive immune response tests for SARS-CoV-2. https://www.fda.gov/medical-devices/coronavirus-disease-2019-covid-19-em.... Accessed 2 December 2021.

[5] Muecksch F, Wise H, Batchelor B, Squires M, Semple E, Richardson C, McGuire J, Clearly S, Furrie E, Greig N, Hay G, Templeton K, Lorenzi JCC, Hatziioannou T, Jenks S, Bieniasz PD. 2021. Longitudinal serological analysis and neutralizing antibody levels in coronavirus disease 2019 convalescent patients. J Infect Dis 223:389–398. doi:10.1093/infdis/jiaa659. - DOI - PMC - PubMed

[6] Muecksch F, Wise H, Batchelor B, Squires M, Semple E, Richardson C, McGuire J, Clearly S, Furrie E, Greig N, Hay G, Templeton K, Lorenzi JCC, Hatziioannou T, Jenks S, Bieniasz PD. 2021. Longitudinal serological analysis and neutralizing antibody levels in coronavirus disease 2019 convalescent patients. J Infect Dis 223:389–398. doi:10.1093/infdis/jiaa659. - DOI - PMC - PubMed

[7] Perez-Saez J, Zaballa M-E, Yerly S, Andrey DO, Meyer B, Eckerle I, Balavoine J-F, Chappuis F, Pittet D, Trono D, Kherad O, Vuilleumier N, Kaiser L, Guessous I, Stringhini S, Azman AS, Specchio-COVID19 Study Group . 2021. Persistence of anti-SARS-CoV-2 antibodies: immunoassay heterogeneity and implications for serosurveillance. Clin Microbiol Infect 27:1695.e7–e1695.e12. doi:10.1016/j.cmi.2021.06.040. - DOI - PMC - PubMed

[8] Perez-Saez J, Zaballa M-E, Yerly S, Andrey DO, Meyer B, Eckerle I, Balavoine J-F, Chappuis F, Pittet D, Trono D, Kherad O, Vuilleumier N, Kaiser L, Guessous I, Stringhini S, Azman AS, Specchio-COVID19 Study Group . 2021. Persistence of anti-SARS-CoV-2 antibodies: immunoassay heterogeneity and implications for serosurveillance. Clin Microbiol Infect 27:1695.e7–e1695.e12. doi:10.1016/j.cmi.2021.06.040. - DOI - PMC - PubMed

[9] Muecksch F, Wise H, Templeton K, Batchelor B, Squires M, McCance K, Jarvis L, Malloy K, Furrie E, Richardson C, MacGuire J, Godber I, Burns A, Mavin S, Zhang F, Schmidt F, Bieniasz P, Jenks S, Hatziioannou T. 2021. Longitudinal variation in SARS-CoV-2 antibody levels and emergence of viral variants: implications for the ability of serological assays to predict immunity. medRxiv doi:10.1101/2021.07.02.21259939. - DOI - PMC - PubMed

[10] Muecksch F, Wise H, Templeton K, Batchelor B, Squires M, McCance K, Jarvis L, Malloy K, Furrie E, Richardson C, MacGuire J, Godber I, Burns A, Mavin S, Zhang F, Schmidt F, Bieniasz P, Jenks S, Hatziioannou T. 2021. Longitudinal variation in SARS-CoV-2 antibody levels and emergence of viral variants: implications for the ability of serological assays to predict immunity. medRxiv doi:10.1101/2021.07.02.21259939. - DOI - PMC - PubMed

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The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization

Affiliations

The Serological Sciences Network (SeroNet) for COVID-19: Depth and Breadth of Serology Assays and Plans for Assay Harmonization

Authors

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Abstract

Conflict of interest statement

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Update of

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