Development of a SARS-CoV-2 Total Antibody Assay and the Dynamics of Antibody Response over Time in Hospitalized and Nonhospitalized Patients with COVID-19

  1. Erik H. Vogelzang
  2. Floris C. Loeff
  3. Ninotska I. L. Derksen
  4. Simone Kruithof
  5. Pleuni Ooijevaar-de Heer
  6. Gerard van Mierlo
  7. Federica Linty
  8. Juk Yee Mok
  9. Wim van Esch
  10. Sanne de Bruin
  11. Alexander P. J. Vlaar
  12. Amsterdam University Medical Center COVID-19 Biobank Study Group
  13. Robert Hemke
  14. Diederik van de Beek
  15. Martijn Beudel
  16. Matthijs C. Brouwer
  17. Bart Geerts
  18. Markus Hollmann
  19. Bennedikt Preckel
  20. Denise Veelo
  21. A. H. (Koos) Zwinderman
  22. Theo Geijtenbeek
  23. Florianne Hafkamp
  24. Diane Bax
  25. Alex Cloherty
  26. Michiel van Agtmael
  27. Marije Bomers
  28. Suzanne Geerlings
  29. Martin P. Grobusch
  30. Vanessa Harris
  31. Sabine M. Hermans
  32. Joppe W. Hovius
  33. Jeaninne Nellen
  34. Edgar Peters
  35. Tom van der Poll
  36. Jan M. Prins
  37. Kim Sigaloff
  38. Cornelis S. Stijnis
  39. Marc van der Valk
  40. Michèle van Vugt
  41. W. Joost Wiersinga
  42. Godelieve Bree
  43. Alexander P. J. Vlaar
  44. Sanne de Bruin
  45. Anne Geke Algera
  46. Frank van Baarle
  47. Lieuwe Bos
  48. Michela Botta
  49. Esther Bulle
  50. Paul Elbers
  51. Lucas Fleuren
  52. Armand Girbes
  53. Laura Hagens
  54. Leo Heunks
  55. Janneke Horn
  56. Mourik van Mourik
  57. Frederique Paulus
  58. Jorinde Raasveld
  59. Marcus Schultz
  60. Marry Smit
  61. Willemke Stilma
  62. Patrick Thoral
  63. Anissa Tsonas
  64. Heder de Vries
  65. Alex Schuurmans
  66. Menno de Jong
  67. Marianna Bugiani
  68. Harm Jan Bogaard
  69. Charlotte Teunissen
  70. Jorg Hamann
  71. Bart Seppen
  72. Maureen Leeuw
  73. Anne J. G. van Oudheusden
  74. Anton G. M. Buiting
  75. Kin Ki Jim
  76. Hans Vrielink
  77. Francis Swaneveld
  78. Gestur Vidarsson
  79. C. Ellen van der Schoot
  80. Peter C. Wever
  81. Wentao Li
  82. Frank van Kuppeveld
  83. Jean-Luc Murk
  84. Berend Jan Bosch
  85. Gerrit-Jan Wolbink
  86. Theo Rispens

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Abstract

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 infections often cause only mild disease that may evoke relatively low Ab titers compared with patients admitted to hospitals. Generally, total Ab bridging assays combine good sensitivity with high specificity. Therefore, we developed sensitive total Ab bridging assays for detection of SARS-CoV-2 Abs to the receptor-binding domain (RBD) and nucleocapsid protein in addition to conventional isotype-specific assays. Ab kinetics was assessed in PCR-confirmed, hospitalized coronavirus disease 2019 (COVID-19) patients (n = 41) and three populations of patients with COVID-19 symptoms not requiring hospital admission: PCR-confirmed convalescent plasmapheresis donors (n = 182), PCR-confirmed hospital care workers (n = 47), and a group of longitudinally sampled symptomatic individuals highly suspect of COVID-19 (n = 14). In nonhospitalized patients, the Ab response to RBD is weaker but follows similar kinetics, as has been observed in hospitalized patients. Across populations, the RBD bridging assay identified most patients correctly as seropositive. In 11/14 of the COVID-19–suspect cases, seroconversion in the RBD bridging assay could be demonstrated before day 12; nucleocapsid protein Abs emerged less consistently. Furthermore, we demonstrated the feasibility of finger-prick sampling for Ab detection against SARS-CoV-2 using these assays. In conclusion, the developed bridging assays reliably detect SARS-CoV-2 Abs in hospitalized and nonhospitalized patients and are therefore well suited to conduct seroprevalence studies.

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  1. Version published to 10.4049/jimmunol.2000767
  2. ScreenIT

    SciScore for 10.1101/2020.06.17.20133793: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementConsent: This study has been conducted in accordance with the ethical principles set out in the declaration of Helsinki and all participants provided written informed consent, if applicable.
    IRB: Approval was obtained from the Medical Ethics Committees from the Academic Medical Center, VU University Medical Center and Elisabeth-TweeSteden Hospital.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableThe average age of participants was 31 years (SD 4.2), 7 (50%) were female and none reported relevant comorbidities.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Development of SARS-CoV-2 RBD and NP based bridging assays: The RBD-mFc and RBD-ST proteins were produced as described by Okba et al.[15] The NP was produced in HEK cells with HAVT20 leader peptide, 10xhis-tag and a BirA-tag as described by Dekkers et al.[24] MaxiSORP microtiter plates (ThermoFisherScientific, US) were coated with 100 µL per well 0.125 µg/mL RBD-mFc or NP in PBS overnight at 4 ° C.
    HEK
    suggested: None
    Software and Algorithms
    SentencesResources
    Data processing and analysis: Statistical analysis were carried out using Graphpad Prism 7.
    Graphpad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    There are some limitations to this study. Further investigation of cross-reactivity for the different coronaviruses may be necessary since the RBD of SARS-CoV-2 may show cross-reactivity with other human coronaviruses (HCoV), and in particular SARS-CoV-1. However, we anticipate that this will not alter our findings due to the limited number of cases with SARS-CoV-1 in Europe. [37] By including a large population of healthy controls the risk of having missed substantial cross-reactivity against other HCoV is also limited; especially since antibodies against HCoV are detected in the majority of the population (>90% for all four HCoV).[38,39] Furthermore, although this study provides an insight into the antibody response in the early phase of infection in non-hospitalized patients, the number of cases is still modest. Our study describes the early antibody kinetics in a population based on probable exposure and not based on symptoms or hospitalized patients who are RT-PCR positive. Yet, this provides an unbiased view of the performance of the developed serological assay and the antibody response in patients with mild SARS-CoV-2 infection. In summary, this study demonstrates the use of a sensitive bridging assay to study seroprevalence in non-hospitalized COVID-19 patients, and provides insight into the early kinetics of the antibody response to SARS-CoV-2 in this population.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2020.06.17.20133793v1 on medRxiv