Abstract
Many severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology tests have proven to be less accurate than expected and do not assess antibody function as neutralizing, correlating with protection from reinfection. A new assay technology measuring the interaction of the purified SARS-CoV-2 spike protein receptor binding domain (RBD) with the extracellular domain of the human angiotensin-converting enzyme 2 (hACE2) receptor detects these important antibodies. The cPass surrogate virus neutralization test (sVNT), compared directly with eight SARS-CoV-2 IgG serology and two live-cell neutralization tests, gives similar or improved accuracy for qualitative delineation between positive and negative individuals in a fast, scalable, and high-throughput assay. The combined data support the cPass sVNT as a tool for highly accurate SARS-CoV-2 immunity surveillance of infected/recovered and/or vaccinated individuals as well as drug and convalescent-phase donor screening. The data also preview a novel application for the cPass sVNT in calibrating the stringency of live-cell neutralization tests and its use in longitudinal testing of recovered and/or vaccinated patients.
Highlights
Many severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology tests have proven to be less accurate than expected and do not assess antibody function as neutralizing, correlating with protection from reinfection
Utilizing 96-well plates coated with the purified extracellular domain of the human angiotensin-converting enzyme 2 receptor and a purified, solubilized, recombinant receptor binding domain (RBD) conjugated to horseradish peroxidase (HRP) (RBD-HRP), the assay capitalizes on the strong interaction between the hACE2 receptor and the RBD coupled with the high immunogenicity of the RBD [8, 17]
For study 1, 68 (45 SARS-CoV-2 PCR presumed positive and 23 prepandemic presumed negative) human serum samples were directly compared across four tests using either RBD (Fig. 2A), nucleocapsid (Fig. 2B), or spike (S1-RBD) (Fig. 2C)-coated enzyme-linked immunosorbent assay (ELISA) plates and the RBD soluble cPass surrogate virus neutralization test (sVNT) (Fig. 2D)
Summary
Many severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology tests have proven to be less accurate than expected and do not assess antibody function as neutralizing, correlating with protection from reinfection. Utilizing 96-well plates coated with the purified extracellular domain of the human angiotensin-converting enzyme 2 (hACE2) receptor and a purified, solubilized, recombinant RBD conjugated to HRP (RBD-HRP), the assay capitalizes on the strong interaction between the hACE2 receptor and the RBD coupled with the high immunogenicity of the RBD [8, 17]. This permits the direct assessment of the inhibitory capacity of immunoglobulins, antibody-based drugs, and compounds that block (or neutralize) this binding event (Fig. 1) [8, 9, 17, 18]
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