tetano
Editor, Senior Moderator
JCI Insight
. 2020 Sep 1;142362.
doi: 10.1172/jci.insight.142362. Online ahead of print.
A simple protein-based surrogate neutralization assay for SARS-CoV-2
Kento T Abe[SUP] 1 [/SUP], Zhijie Li[SUP] 2 [/SUP], Reuben Samson[SUP] 1 [/SUP], Payman Samavarchi-Tehrani[SUP] 1 [/SUP], Emelissa J Valcourt[SUP] 3 [/SUP], Heidi Wood[SUP] 3 [/SUP], Patrick Budylowski[SUP] 4 [/SUP], Alan P Dupuis Ii[SUP] 5 [/SUP], Roxie C Girardin[SUP] 5 [/SUP], Bhavisha Rathod[SUP] 1 [/SUP], Jenny Wang[SUP] 1 [/SUP], Miriam Barrios-Rodiles[SUP] 1 [/SUP], Karen Colwill[SUP] 1 [/SUP], Allison McGeer[SUP] 6 [/SUP], Samira Mubareka[SUP] 7 [/SUP], Jennifer L Gommerman[SUP] 4 [/SUP], Yves Durocher[SUP] 8 [/SUP], Mario Ostrowski[SUP] 4 [/SUP], Kathleen A McDonough[SUP] 5 [/SUP], Michael A Drebot[SUP] 3 [/SUP], Steven J Drews[SUP] 9 [/SUP], James M Rini[SUP] 10 [/SUP], Anne-Claude Gingras[SUP] 11 [/SUP]
Affiliations
Abstract
Most of the patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mount a humoral immune response to the virus within a few weeks of infection, but the duration of this response and how it correlates with clinical outcomes has not been completely characterized. Of particular importance is the identification of immune correlates of infection that would support public health decision-making on treatment approaches, vaccination strategies, and convalescent plasma therapy. While ELISA-based assays to detect and quantitate antibodies to SARS-CoV-2 in patient samples have been developed, the detection of neutralizing antibodies typically requires more demanding cell-based viral assays. Here, we present a safe and efficient protein-based assay for the detection of serum and plasma antibodies that block the interaction of the SARS-CoV-2 spike protein receptor binding domain (RBD) with its receptor, angiotensin converting-enzyme 2 (ACE2). The assay serves as a surrogate neutralization assay and is performed on the same platform and in parallel with an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against the RBD, enabling a direct comparison. The results obtained with our assay correlate with those of two viral based assays, a plaque reduction neutralization test (PRNT) that uses live SARS-CoV-2 virus, and a spike pseudotyped viral-vector-based assay.
Keywords: Adaptive immunity; Antigen; Immunoglobulins; Infectious disease.
. 2020 Sep 1;142362.
doi: 10.1172/jci.insight.142362. Online ahead of print.
A simple protein-based surrogate neutralization assay for SARS-CoV-2
Kento T Abe[SUP] 1 [/SUP], Zhijie Li[SUP] 2 [/SUP], Reuben Samson[SUP] 1 [/SUP], Payman Samavarchi-Tehrani[SUP] 1 [/SUP], Emelissa J Valcourt[SUP] 3 [/SUP], Heidi Wood[SUP] 3 [/SUP], Patrick Budylowski[SUP] 4 [/SUP], Alan P Dupuis Ii[SUP] 5 [/SUP], Roxie C Girardin[SUP] 5 [/SUP], Bhavisha Rathod[SUP] 1 [/SUP], Jenny Wang[SUP] 1 [/SUP], Miriam Barrios-Rodiles[SUP] 1 [/SUP], Karen Colwill[SUP] 1 [/SUP], Allison McGeer[SUP] 6 [/SUP], Samira Mubareka[SUP] 7 [/SUP], Jennifer L Gommerman[SUP] 4 [/SUP], Yves Durocher[SUP] 8 [/SUP], Mario Ostrowski[SUP] 4 [/SUP], Kathleen A McDonough[SUP] 5 [/SUP], Michael A Drebot[SUP] 3 [/SUP], Steven J Drews[SUP] 9 [/SUP], James M Rini[SUP] 10 [/SUP], Anne-Claude Gingras[SUP] 11 [/SUP]
Affiliations
- PMID: 32870820
- DOI: 10.1172/jci.insight.142362
Abstract
Most of the patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mount a humoral immune response to the virus within a few weeks of infection, but the duration of this response and how it correlates with clinical outcomes has not been completely characterized. Of particular importance is the identification of immune correlates of infection that would support public health decision-making on treatment approaches, vaccination strategies, and convalescent plasma therapy. While ELISA-based assays to detect and quantitate antibodies to SARS-CoV-2 in patient samples have been developed, the detection of neutralizing antibodies typically requires more demanding cell-based viral assays. Here, we present a safe and efficient protein-based assay for the detection of serum and plasma antibodies that block the interaction of the SARS-CoV-2 spike protein receptor binding domain (RBD) with its receptor, angiotensin converting-enzyme 2 (ACE2). The assay serves as a surrogate neutralization assay and is performed on the same platform and in parallel with an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against the RBD, enabling a direct comparison. The results obtained with our assay correlate with those of two viral based assays, a plaque reduction neutralization test (PRNT) that uses live SARS-CoV-2 virus, and a spike pseudotyped viral-vector-based assay.
Keywords: Adaptive immunity; Antigen; Immunoglobulins; Infectious disease.