1. A surrogate cell-based SARS-CoV-2 spike blocking assay.
- Author
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Schuh W, Baus L, Steinmetz T, Schulz SR, Weckwerth L, Roth E, Hauke M, Krause S, Morhart P, Rauh M, Hoffmann M, Vesper N, Reth M, Schneider H, Jäck HM, and Mielenz D
- Subjects
- Antibodies, Blocking immunology, Antibodies, Neutralizing immunology, COVID-19 immunology, Humans, SARS-CoV-2, Spike Glycoprotein, Coronavirus immunology, Antibodies, Blocking blood, Antibodies, Neutralizing blood, Antibodies, Viral analysis, COVID-19 diagnosis, COVID-19 Serological Testing methods, Flow Cytometry methods
- 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., (© 2021 The Authors. European Journal of Immunology published by Wiley-VCH GmbH.)
- Published
- 2021
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