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

Ji Woong Kim; Ah Hyun Cho; Ha Gyeong Shin; Sung Hoon Jang; Su Yeon Cho; Ye Rim Lee; Sukmook Lee

doi:10.3390/v15010174

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

Viruses. 2023 Jan 6;15(1):174. doi: 10.3390/v15010174.

Authors

Ji Woong Kim¹, Ah Hyun Cho², Ha Gyeong Shin², Sung Hoon Jang¹, Su Yeon Cho², Ye Rim Lee², Sukmook Lee^{1

2

3}

Affiliations

¹ Department of Applied Chemistry, Kookmin University, Seoul 02707, Republic of Korea.
² Biopharmaceutical Chemistry Major, School of Applied Chemistry, Kookmin University, Seoul 02707, Republic of Korea.
³ Antibody Research Institute, Kookmin University, Seoul 02707, Republic of Korea.

Abstract

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.

Keywords: SARS-CoV-2; phage display; sandwich immunoassay; spike protein.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Monoclonal* / genetics
Antibodies, Neutralizing
Antibodies, Viral
Bacteriophages*
COVID-19
Humans
SARS-CoV-2* / genetics
Single-Chain Antibodies* / genetics
Spike Glycoprotein, Coronavirus

Substances

Antibodies, Monoclonal
Antibodies, Neutralizing
Antibodies, Viral
Single-Chain Antibodies
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2

Supplementary concepts

SARS-CoV-2 variants

Grants and funding

This research was supported by the Bio & Medical Technology Development Program of the National Research Foundation of Korea (grant numbers: NRF-2019M3E5D5065844 and NRF-2020M3A9I2107093) and the Korea Health Technology R&D Project of the Korea Health Industry Development Institute (grant number: HI22C0360) funded by the Korean government.