Abstract
The establishment of an approach for detecting the anti-severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-receptor-binding domain (RBD) neutralizing antibodies (nAbs) by a safe, easy, and rapid technique without requiring the use of live viruses is essential for facing the coronavirus disease 2019 (COVID-19) pandemic. Depending on competitive enzyme-linked immunosorbent assay (ELISA) methodology, the current study assay was designed to simulate the virus-host interaction using purified SARS-COV-2-RBD from the spike protein and the host cell receptor human angiotensin-converting enzyme 2 protein. The performance of this in-house neutralizing ELISA assay was validated using freshly prepared standards with different known concentrations of the assay. In this regard, a cohort of 50 serum samples from convalescent COVID-19 individuals with different disease severity at different time points post-recovery and a cohort of 50 serum samples from healthy individuals were processed by the in-house developed assay for detecting SARS-CoV-2 nAbs, in comparison with a commercial total anti-SARS-CoV-2 IgG antibody assay as a gold standard. The assay obtained a sensitivity of 88% (95% CI: 75.69-95.47) and a specificity of 92% (95% CI: 80.77- 97.78%). A negative strong correlation was demonstrated in the standard curve between the optical density absorbance and log concentration of the nAbs with a statistical measure of r2 (coefficient of determination) = 0.9539. The SARS-COV-2-RBD neutralizing ELISA assay serves as a high throughput qualitative and quantitative tool that can be applied in most laboratory settings without special biosafety requirements to detect anti-RBD nAbs for seroprevalence, pre-clinical, and clinical evaluation of COVID-19 vaccines efficiency and the rapid selection of convalescent plasma donors for the treatment of COVID-19 patients.
Keywords: COVID-19; SARS-COV-2-RBD neutralizing ELISA assay; nAbs.
【저자키워드】 COVID-19, SARS-COV-2-RBD neutralizing ELISA assay, nAbs., 【초록키워드】 Treatment, coronavirus disease, viruses, convalescent plasma, neutralizing antibody, SARS-CoV-2, Coronavirus disease 2019, COVID-19 vaccine, pandemic, Neutralizing antibodies, antibody, disease severity, Seroprevalence, ELISA, Spike protein, Laboratory, biosafety, Protein, sensitivity, specificity, enzyme-linked immunosorbent assay, Cohort, COVID-19 vaccines, clinical evaluation, anti-SARS-CoV-2 IgG, RBD, gold, optical, Neutralizing, methodology, respiratory, convalescent, correlation, Quantitative, COVID-19 patients, ELISA assay, NAb, Donor, Angiotensin-converting enzyme, Coronavirus-2, Interaction, Concentration, angiotensin, standard curve, Efficiency, anti-RBD, Safe, host cell, optical density, acute respiratory syndrome, 95% CI, acute respiratory syndrome coronavirus, serum samples, house, healthy individuals, different time points, gold standard, individual, NAbs, domain, human Angiotensin-converting enzyme, treatment of COVID-19 patients, host cell receptor, serum sample, live viruses, approach, statistical, SARS-CoV-2 nAbs, enzyme-linked immunosorbent, Coefficient of determination, log, detect, the spike protein, demonstrated, purified, different time point, processed, healthy individual, absorbance, live virus, 【제목키워드】 ELISA, validation, application, SARS-CoV-2 RBD,