Development and Characterization of Phage Display-Derived Monoclonal Antibodies to the S2 Domain of Spike Proteins of Wild-Type SARS-CoV-2 and Multiple Variants

The rapid emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has resulted in the ongoing global coronavirus disease 2019 (COVID-19) pandemic. Thus, the rapid development of a platform to detect a broad range of SARS-CoV-2 variants is essential for successful COVID...

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Bibliographic Details
Published in:Viruses 2023-01, Vol.15 (1), p.174
Main Authors: Kim, Ji Woong, Cho, Ah Hyun, Shin, Ha Gyeong, Jang, Sung Hoon, Cho, Su Yeon, Lee, Ye Rim, Lee, Sukmook
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - genetics
Antibodies, Neutralizing
Antibodies, Viral
Antibody libraries
Antigens
Bacteriophages
Cloning
Coronaviruses
COVID-19
Disease transmission
Enzymes
Humans
Immunoassay
Infections
Monoclonal antibodies
Mutation
Phage display
Proteins
sandwich immunoassay
SARS-CoV-2
SARS-CoV-2 - genetics
Severe acute respiratory syndrome coronavirus 2
Single-Chain Antibodies - genetics
Spike Glycoprotein, Coronavirus
Spike protein
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Summary:The rapid emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has resulted in the ongoing global coronavirus disease 2019 (COVID-19) pandemic. Thus, the rapid development of a platform to detect a broad range of SARS-CoV-2 variants is essential for successful COVID-19 management. In this study, four SARS-CoV-2 spike protein-specific single-chain variable fragments (scFvs) were isolated from a synthetic antibody library using phage display technology. Following the conversion of these scFvs into monoclonal antibodies (mAbs) (K104.1-K104.4) and production and purification of the mAbs, the antibody pair (K104.1 and K104.2) that exhibited the highest binding affinity (K104.1 and K104.2, 1.3 nM and 1.9 nM) was selected. Biochemical analyses revealed that this antibody pair specifically bound to different sites on the S2 subunit of the spike protein. Furthermore, we developed a highly sensitive sandwich immunoassay using this antibody pair that accurately and quantitatively detected the spike proteins of wild-type SARS-CoV-2 and multiple variants, including Alpha, Beta, Gamma, Delta, Kappa, and Omicron, in the picomolar range. Conclusively, the novel phage display-derived mAbs we have developed may be useful for the rapid and efficient detection of the fast-evolving SARS-CoV-2.
ISSN:1999-4915
1999-4915
DOI:10.3390/v15010174