A rapid, point-of-care red blood cell agglutination assay detecting antibodies against SARS-CoV-2
AuthorKruse, Robert L
Gehrie, Eric A
Amukele, Timothy K
Tobian, Aaron A R
Mostafa, Heba H
Wang, Zack Z
JournalBiochemical and Biophysical Research Communications
MetadataShow full item record
AbstractThe COVID-19 pandemic has caused significant morbidity and mortality. There is an urgent need for serological tests to detect antibodies against SARS-CoV-2, which could be used to assess past infection, evaluate responses to vaccines in development, and determine individuals who may be protected from future infection. Current serological tests developed for SARS-CoV-2 rely on traditional technologies such as enzyme-linked immunosorbent assays (ELISA) and lateral flow assays, which have not scaled to meet the demand of hundreds of millions of antibody tests so far. Herein, we present an alternative method of antibody testing that depends on one protein reagent being added to patient serum/plasma or whole blood with direct, visual readout. Two novel fusion proteins, RBD-2E8 and B6–CH1-RBD, were designed to bind red blood cells (RBCs) via a single-chain variable fragment (scFv), thereby displaying the receptor-binding domain (RBD) of SARS-CoV-2 spike protein on the surface of RBCs. Mixing mammalian-derived RBD-2E8 and B6–CH1-RBD with convalescent COVID-19 patient serum and RBCs led to visible hemagglutination, indicating the presence of antibodies against SARS-CoV-2 RBD. B6–CH1-RBD made in bacteria was not as effective in inducing agglutination, indicating better recognition of RBD epitopes from mammalian cells. Given that our hemagglutination test uses methods routinely used in hospital clinical labs across the world for blood typing, we anticipate the test can be rapidly deployed at minimal cost. We anticipate our hemagglutination assay may find extensive use in low-resource settings for detecting SARS-CoV-2 antibodies. © 2021 Elsevier Inc.
SponsorsThis work was funded by the Johns Hopkins Department of Pathology Fred and Janet Sanfilippo Research Award (R.L.K), and partially supported by the National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Disease [grant R01DK106109] (Z.Z.W).
Rights/TermsCopyright © 2021 Elsevier Inc. All rights reserved.
Identifier to cite or link to this itemhttp://hdl.handle.net/10713/15106