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Surrogate Virus Neutralisation Test Based on Nanoluciferase-Tagged Antigens to Quantify Inhibitory Antibodies against SARS-CoV-2 and Characterise Omicron-Specific Reactivity in a Vaccination Cohort

ORCID
0000-0002-3064-4152
Affiliation
Institute of Virology, Department of Molecular Virology, University of Muenster, Von-Esmarch-Str. 56, D-48149 Muenster, Germany;(M.S.);(E.R.H.);(S.L.)
Schoefbaenker, Michael;
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Neddermeyer, Rieke;
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Guenther, Theresa;
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Mueller, Marlin M.;
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Romberg, Marie-Luise;
ORCID
0000-0002-2745-5748
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Classen, Nica;
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Hennies, Marc T.;
ORCID
0000-0003-3145-5887
Affiliation
Institute of Virology, Department of Molecular Virology, University of Muenster, Von-Esmarch-Str. 56, D-48149 Muenster, Germany;(M.S.);(E.R.H.);(S.L.)
Hrincius, Eike R.;
ORCID
0000-0003-4490-3052
Affiliation
Institute of Virology, Department of Molecular Virology, University of Muenster, Von-Esmarch-Str. 56, D-48149 Muenster, Germany;(M.S.);(E.R.H.);(S.L.)
Ludwig, Stephan;
ORCID
0000-0003-2992-1471
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Kuehn, Joachim E.;
ORCID
0000-0002-5570-1897
Affiliation
Institute of Virology, Department of Clinical Virology, University of Muenster, Von-Stauffenberg-Str. 36, D-48151 Muenster, Germany;(R.N.);(T.G.);(M.M.M.);(M.-L.R.);(N.C.);(M.T.H.);(J.E.K.)
Lorentzen, Eva U.

Virus-specific antibodies are crucial for protective immunity against SARS-CoV-2. Assessing functional antibodies through conventional or pseudotyped virus neutralisation tests (pVNT) requires high biosafety levels. Alternatively, the virus-free surrogate virus neutralisation test (sVNT) quantifies antibodies interfering with spike binding to angiotensin-converting enzyme 2. We evaluated secreted nanoluciferase-tagged spike protein fragments as diagnostic antigens in the sVNT in a vaccination cohort. Initially, spike fragments were tested in a capture enzyme immunoassay (EIA), identifying the receptor binding domain (RBD) as the optimal diagnostic antigen. The sensitivity of the in-house sVNT applying the nanoluciferase-labelled RBD equalled or surpassed that of a commercial sVNT ( cPass , GenScript Diagnostics) and an in-house pVNT four weeks after the first vaccination (98% vs. 94% and 72%, respectively), reaching 100% in all assays four weeks after the second and third vaccinations. When testing serum reactivity with Omicron BA.1 spike, the sVNT and pVNT displayed superior discrimination between wild-type- and variant-specific serum reactivity compared to a capture EIA. This was most pronounced after the first and second vaccinations, with the third vaccination resulting in robust, cross-reactive BA.1 construct detection. In conclusion, utilising nanoluciferase-labelled antigens permits the quantification of SARS-CoV-2-specific inhibitory antibodies. Designed as flexible modular systems, the assays can be readily adjusted for monitoring vaccine efficacy.

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Editor:
Venketaraman, Vishwanath
Published in:
Vaccines  
Vol. 11, No. 12 (08.12.2023)
Volume:
11
No.:
12
Date Issued:
08.12.2023
DOI:
10.3390/vaccines11121832
Language:
English
Type of Resource:
Text
Keywords:
  1. SARS-CoV-2
    Typ
    Schlagwort
  1. Omicron BA.1
    Typ
    Schlagwort
  1. surrogate virus neutralisation test
    Typ
    Schlagwort
  1. pseudotyped virus neutralisation test
    Typ
    Schlagwort
  1. antibody quantification
    Typ
    Schlagwort
  1. nanoluciferase
    Typ
    Schlagwort
  1. vaccination study
    Typ
    Schlagwort
DDC:
610 Medizin und Gesundheit

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