Assessment of specific human antibodies against SARS-CoV-2 receptor binding domain by rapid in-house ELISA

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Hussein N.A.; Ali E.A.A.; El-Hakim A.E.; Tabll A.A.; El-Shershaby A.; Salamony A.; Shaheen M.N.F.; Ali I.; Elshall M.; Shahein Y.E.

Journal: Human Antibodies

Publisher: IOS Press BV

Publication Date: 12 May 2022

Volume / Issue: Volume 30 / Issue 2

Pages: 105–115

ISSN: 10932607

DOI: 10.3233/HAB-220003

Scopus: View on Scopus

PubMed: 35431235

Document Type: Article

Access: All Open Access; Bronze Open Access

Authors and Affiliations

Hussein N.A., Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt; Ali E.A.A., Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt; El-Hakim A.E., Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt; Tabll A.A., Microbial Biotechnology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt, Egypt Center for Research and Regenerative Medicine, Cairo, Egypt; El-Shershaby A., Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt; Salamony A., Egypt Center for Research and Regenerative Medicine, Cairo, Egypt; Shaheen M.N.F., Environmental Virology Laboratory, Water Pollution Research Department, Environmental and Climate Change Research Institute, National Research Centre, Dokki, Cairo, Egypt; Ali I., Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt; Elshall M., Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt; Shahein Y.E., Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, Egypt

Abstract

BACKGROUND: The recently emerged SARS-CoV-2 caused a global pandemic since the last two years. The urgent need to control the spread of the virus and rapid application of the suitable health measures raised the importance of available, rapid, and accurate diagnostic approaches. OBJECTIVE: The purpose of this study is to describe a rapid in-house optimized ELISA based on the expression of the receptor binding domain (RBD) of the SARS-CoV-2 spike protein in a prokaryotic system. METHODS: We show the expression of the 30 kDa recombinant SARS-CoV-2 RBD-6×His in four different E. coli strains (at 28∘C using 0.25mM IPTG) including the expression strain E. coli BL21 (DE3) Rosetta Gami. SARS-CoV-2 rRBD-6×His protein was purified, refolded, and used as an antigen coat to assess antibody response in human sera against SARS-CoV-2 infection. RESULTS: The assessment was carried out using a total of 155 human sero-positive and negative SARS-CoV-2 antibodies. The ELISA showed 69.5% sensitivity, 88% specificity, 78.5% agreement, a positive predictive value (PPV) of 92.3%, and a negative predictive value of 56.5%. Moreover, the optical density (OD) values of positive samples significantly correlated with the commercial kit titers. CONCLUSIONS: Specific human antibodies against SARS-CoV-2 spike protein were detected by rapid in-house ELISA in sera of human COVID-19-infected patients. The availability of this in-house ELISA protocol would be valuable for various diagnostic and epidemiological applications, particularly in developing countries. Future studies are planned for the use of the generated SARS-CoV-2 rRBD-6×His protein in vaccine development and other diagnostic applications. © 2022 - IOS Press. All rights reserved.

Keywords

COVID-19; diagnosis; ELISA; human sera; RBD; SARS-CoV-2; Antibodies, Viral; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Humans; Spike Glycoprotein, Coronavirus; antibody; isopropyl thiogalactoside; coronavirus spike glycoprotein; spike protein, SARS-CoV-2; virus antibody; antibody response; Article; coronavirus disease 2019; enzyme linked immunosorbent assay; expression vector; gene expression; gene overexpression; limit of detection; nonhuman; nucleotide sequence; plasmid; polyacrylamide gel electrophoresis; predictive value; protein expression; protein purification; protein renaturation; receptor binding; Sanger sequencing; sensitivity and specificity; Severe acute respiratory syndrome coronavirus 2; genetics; human; procedures

Citation Information

Scopus Citations: 7

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