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A surrogate cell-based SARS-CoV-2 spike blocking assay.
Schuh, Wolfgang; Baus, Lena; Steinmetz, Tobit; Schulz, Sebastian R; Weckwerth, Leonie; Roth, Edith; Hauke, Manuela; Krause, Sara; Morhart, Patrick; Rauh, Manfred; Hoffmann, Markus; Vesper, Niklas; Reth, Michael; Schneider, Holm; Jäck, Hans-Martin; Mielenz, Dirk.
  • Schuh W; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Baus L; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Steinmetz T; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Schulz SR; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Weckwerth L; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Roth E; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Hauke M; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Krause S; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Morhart P; Division of Molecular Pediatrics, Department of Pediatrics, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Rauh M; Division of Molecular Pediatrics, Department of Pediatrics, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Hoffmann M; Infection Biology Unit, German Primate Center-Leibniz Institute for Primate Research, Göttingen, Germany.
  • Vesper N; Faculty of Biology and Psychology, University of Göttingen, Göttingen, Germany.
  • Reth M; Institute of Biology III (Molecular Immunology), University of Freiburg, Freiburg, Germany.
  • Schneider H; Institute of Biology III (Molecular Immunology), University of Freiburg, Freiburg, Germany.
  • Jäck HM; Division of Molecular Pediatrics, Department of Pediatrics, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
  • Mielenz D; Division of Molecular Immunology, Department of Internal Medicine 3, Nikolaus-Fiebiger-Zentrum, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, Erlangen, Germany.
Eur J Immunol ; 51(11): 2665-2676, 2021 11.
Article in English | MEDLINE | ID: covidwho-1482126
ABSTRACT
To monitor infection by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and successful vaccination against coronavirus disease 2019 (COVID-19), the kinetics of neutralizing or blocking anti-SARS-CoV-2 antibody titers need to be assessed. Here, we report the development of a quick and inexpensive surrogate SARS-CoV-2 blocking assay (SUBA) using immobilized recombinant human angiotensin-converting enzyme 2 (hACE2) and human cells expressing the native form of surface SARS-CoV-2 spike protein. Spike protein-expressing cells bound to hACE2 in the absence or presence of blocking antibodies were quantified by measuring the optical density of cell-associated crystal violet in a spectrophotometer. The advantages are that SUBA is a fast and inexpensive assay, which does not require biosafety level 2- or 3-approved laboratories. Most importantly, SUBA detects blocking antibodies against the native trimeric cell-bound SARS-CoV-2 spike protein and can be rapidly adjusted to quickly pre-screen already approved therapeutic antibodies or sera from vaccinated individuals for their ACE2 blocking activities against any emerging SARS-CoV-2 variants.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Antibodies, Blocking / Antibodies, Neutralizing / Flow Cytometry / COVID-19 Serological Testing / COVID-19 / Antibodies, Viral Topics: Vaccines / Variants Limits: Humans Language: English Journal: Eur J Immunol Year: 2021 Document Type: Article Affiliation country: Eji.202149302

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Antibodies, Blocking / Antibodies, Neutralizing / Flow Cytometry / COVID-19 Serological Testing / COVID-19 / Antibodies, Viral Topics: Vaccines / Variants Limits: Humans Language: English Journal: Eur J Immunol Year: 2021 Document Type: Article Affiliation country: Eji.202149302