tetano
Editor, Senior Moderator
Vaccines (Basel)
. 2020 Jul 15;8(3):E386.
doi: 10.3390/vaccines8030386.
Rapid Quantification of SARS-CoV-2-Neutralizing Antibodies Using Propagation-Defective Vesicular Stomatitis Virus Pseudotypes
Ferdinand Zettl[SUP] 1 [/SUP], Toni Luise Meister[SUP] 2 [/SUP], Tanja Vollmer[SUP] 3 [/SUP], Bastian Fischer[SUP] 3 [/SUP], J?rg Steinmann[SUP] 4 5 [/SUP], Adalbert Krawczyk[SUP] 6 7 [/SUP], Philip V'kovski[SUP] 1 8 [/SUP], Daniel Todt[SUP] 2 [/SUP], Eike Steinmann[SUP] 2 [/SUP], Stephanie Pfaender[SUP] 2 [/SUP], Gert Zimmer[SUP] 1 8 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2, a new member of the genus Betacoronavirus, is a pandemic virus, which has caused numerous fatalities, particularly in the elderly and persons with underlying morbidities. At present, there are no approved vaccines nor antiviral therapies available. The detection and quantification of SARS-CoV-2-neutralizing antibodies plays a crucial role in the assessment of the immune status of convalescent COVID-19 patients, evaluation of recombinant therapeutic antibodies, and the evaluation of novel vaccines. To detect SARS-CoV-2-neutralizing antibodies, classically, a virus-neutralization test has to be performed at biosafety level 3, considerably limiting the general use of this test. In the present work, a biosafety level 1 pseudotype virus assay based on a propagation-incompetent vesicular stomatitis virus (VSV) has been used to determine the neutralizing antibody titers in convalescent COVID-19 patients. The neutralization titers in serum of two independently analyzed patient cohorts were available within 18 h and correlated well with those obtained with a classical SARS-CoV-2 neutralization test (Pearson correlation coefficients of r = 0.929 and r = 0.939, respectively). Most convalescent COVID-19 patients had only low titers of neutralizing antibodies (ND50 < 320). The sera of convalescent COVID-19 patients also neutralized pseudotype virus displaying the SARS-CoV-1 spike protein on their surface, which is homologous to the SARS-CoV-2 spike protein. In summary, we report a robust virus-neutralization assay, which can be used at low biosafety level 1 to rapidly quantify SARS-CoV-2-neutralizing antibodies in convalescent COVID-19 patients and vaccinated individuals.
Keywords: SARS-CoV-2; neutralizing antibodies; passive immunization; pseudovirus; vaccine.
. 2020 Jul 15;8(3):E386.
doi: 10.3390/vaccines8030386.
Rapid Quantification of SARS-CoV-2-Neutralizing Antibodies Using Propagation-Defective Vesicular Stomatitis Virus Pseudotypes
Ferdinand Zettl[SUP] 1 [/SUP], Toni Luise Meister[SUP] 2 [/SUP], Tanja Vollmer[SUP] 3 [/SUP], Bastian Fischer[SUP] 3 [/SUP], J?rg Steinmann[SUP] 4 5 [/SUP], Adalbert Krawczyk[SUP] 6 7 [/SUP], Philip V'kovski[SUP] 1 8 [/SUP], Daniel Todt[SUP] 2 [/SUP], Eike Steinmann[SUP] 2 [/SUP], Stephanie Pfaender[SUP] 2 [/SUP], Gert Zimmer[SUP] 1 8 [/SUP]
Affiliations
- PMID: 32679691
- DOI: 10.3390/vaccines8030386
Abstract
Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2, a new member of the genus Betacoronavirus, is a pandemic virus, which has caused numerous fatalities, particularly in the elderly and persons with underlying morbidities. At present, there are no approved vaccines nor antiviral therapies available. The detection and quantification of SARS-CoV-2-neutralizing antibodies plays a crucial role in the assessment of the immune status of convalescent COVID-19 patients, evaluation of recombinant therapeutic antibodies, and the evaluation of novel vaccines. To detect SARS-CoV-2-neutralizing antibodies, classically, a virus-neutralization test has to be performed at biosafety level 3, considerably limiting the general use of this test. In the present work, a biosafety level 1 pseudotype virus assay based on a propagation-incompetent vesicular stomatitis virus (VSV) has been used to determine the neutralizing antibody titers in convalescent COVID-19 patients. The neutralization titers in serum of two independently analyzed patient cohorts were available within 18 h and correlated well with those obtained with a classical SARS-CoV-2 neutralization test (Pearson correlation coefficients of r = 0.929 and r = 0.939, respectively). Most convalescent COVID-19 patients had only low titers of neutralizing antibodies (ND50 < 320). The sera of convalescent COVID-19 patients also neutralized pseudotype virus displaying the SARS-CoV-1 spike protein on their surface, which is homologous to the SARS-CoV-2 spike protein. In summary, we report a robust virus-neutralization assay, which can be used at low biosafety level 1 to rapidly quantify SARS-CoV-2-neutralizing antibodies in convalescent COVID-19 patients and vaccinated individuals.
Keywords: SARS-CoV-2; neutralizing antibodies; passive immunization; pseudovirus; vaccine.