Abstract

Better diagnostic tools are needed to combat the ongoing COVID-19 pandemic. Here, to meet this urgent demand, we report a homogeneous immunoassay to detect IgG antibodies against SARS-CoV-2. This serological assay, called SATiN, is based on a tri-part Nanoluciferase (tNLuc) approach, in which the spike protein of SARS-CoV-2 and protein G, fused respectively to two different tNLuc tags, are used as antibody probes. Target engagement of the probes allows reconstitution of a functional luciferase in the presence of the third tNLuc component. The assay is performed directly in the liquid phase of patient sera and enables rapid, quantitative and low-cost detection. We show that SATiN has a similar sensitivity to ELISA, and its readouts are consistent with various neutralizing antibody assays. This proof-of-principle study suggests potential applications in diagnostics, as well as disease and vaccination management.

Serological tests are important diagnostic and disease surveillance tools for addressing the COVID-19 pandemic. Here, the authors present a tri-part Nanoluciferase based assay to detect antibodies against SARS-CoV-2.

Details

Title
A homogeneous split-luciferase assay for rapid and sensitive detection of anti-SARS CoV-2 antibodies
Author
Yao Zhong 1 ; Drecun Luka 2 ; Aboualizadeh Farzaneh 1 ; Kim Sun Jin 3 ; Li, Zhijie 4   VIAFID ORCID Logo  ; Wood, Heidi 5 ; Valcourt, Emelissa J 5 ; Manguiat Kathy 5 ; Plenderleith Simon 6 ; Yip, Lily 6   VIAFID ORCID Logo  ; Li Xinliu 7 ; Zhong, Zoe 7 ; Yue Feng Yun 8 ; Closas Tatiana 9 ; Snider, Jamie 1 ; Tomic Jelena 1 ; Drews, Steven J 10 ; Drebot, Michael A 5 ; McGeer, Allison 7 ; Ostrowski, Mario 8 ; Mubareka Samira 11 ; Rini, James M 12 ; Owen, Shawn 13   VIAFID ORCID Logo  ; Stagljar Igor 14   VIAFID ORCID Logo 

1  University of Toronto, Donnelly Centre, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Biochemistry, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
2  University of Toronto, Donnelly Centre, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Molecular Genetics, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
3  University of Utah, Department of Pharmaceutics and Pharmaceutical Chemistry, Salt Lake City, USA (GRID:grid.223827.e) (ISNI:0000 0001 2193 0096) 
4  University of Toronto, Department of Molecular Genetics, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
5  Zoonotic Diseases and Special Pathogens Division, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Canada (GRID:grid.415368.d) (ISNI:0000 0001 0805 4386) 
6  Sunnybrook Research Institute, Biological Sciences, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
7  Department of Microbiology, Mount Sinai Hospital, Toronto, Canada (GRID:grid.416166.2) (ISNI:0000 0004 0473 9881) 
8  University of Toronto, Department of Laboratory Medicine and Pathobiology, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
9  Canadian Blood Services, Vancouver, Canada (GRID:grid.423370.1) (ISNI:0000 0001 0285 1288) 
10  Canadian Blood Services, Edmonton, Canada (GRID:grid.423370.1) (ISNI:0000 0001 0285 1288); University of Alberta, Department of Laboratory Medicine & Pathology, Edmonton, Canada (GRID:grid.17089.37) 
11  Sunnybrook Research Institute, Biological Sciences, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Laboratory Medicine and Pathobiology, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
12  University of Toronto, Department of Biochemistry, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Molecular Genetics, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938) 
13  University of Utah, Department of Pharmaceutics and Pharmaceutical Chemistry, Salt Lake City, USA (GRID:grid.223827.e) (ISNI:0000 0001 2193 0096); University of Utah, Department of Biomedical Engineering, Salt Lake City, USA (GRID:grid.223827.e) (ISNI:0000 0001 2193 0096); University of Utah, Department of Medicinal Chemistry, Salt Lake City, USA (GRID:grid.223827.e) (ISNI:0000 0001 2193 0096) 
14  University of Toronto, Donnelly Centre, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Biochemistry, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); University of Toronto, Department of Molecular Genetics, Toronto, Canada (GRID:grid.17063.33) (ISNI:0000 0001 2157 2938); Mediterranean Institute for Life Sciences, Split, Croatia (GRID:grid.482535.d) (ISNI:0000 0004 4663 8413) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-021-22102-6
ProQuest document ID
2503525504
Copyright
© The Author(s) 2021. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.