A rapid, point-of-care red blood cell agglutination assay detecting antibodies against SARS-CoV-2.

Autor: Kruse RL; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: robert.kruse@aya.yale.edu., Huang Y; Department of Medicine, University of Maryland Medical Center Midtown Campus, Baltimore, MD, USA; Division of Gastroenterology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Smetana H; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Gehrie EA; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Amukele TK; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Tobian AAR; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Mostafa HH; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Wang ZZ; Division of Hematology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: zack.wang@jhu.edu.
Jazyk: angličtina
Zdroj: Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2021 May 14; Vol. 553, pp. 165-171. Date of Electronic Publication: 2021 Mar 15.
DOI: 10.1016/j.bbrc.2021.03.016
Abstrakt: The 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.
Competing Interests: Declaration of competing interest R.L.K. is an inventor on a provisional patent application related to the work described in the manuscript. All other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2021 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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