Chatikorn Boonkrai, Thomas S. Cotrone, Watchadaporn Chaisuriyong, Terapong Tantawichien, Usa Thisyakorn, Stefan Fernandez, Taweewun Hunsawong, Matthew Reed, Tossapon Wongtangprasert, Thittaya Audomsun, Tanapati Phakham, Chadaporn Attakitbancha, Pijitra Saelao, Dorota Focht, Raymond Kimbung, Martin Welin, Aijaz Ahmad Malik, Trairak Pisitkun, Nattachai Srisawat
Abstract
The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently the biggest healthcare issue worldwide. This study aimed to develop a monoclonal antibody against SARS-CoV-2 from B cells of recovered COVID-19 patients, which might have beneficial therapeutic purposes for COVID-19 patients. We successfully generated human monoclonal antibodies (hmAbs) against the receptor binding domain (RBD) protein of SARS-CoV-2 using developed hybridoma technology.
Introduction
Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has rapidly spread causing a worldwide pandemic. The SARS-CoV-2 is an RNA virus with the characteristic of multiple spike glycoproteins on its envelope. The receptor-binding domain (RBD) on the spike proteins binds specifically with the cellular receptor angiotensin-converting enzyme 2 (ACE2) of its host cells, resulting in a fusion cascade and virus entry.
Materials and Methods:
Ethical statement
This project was approved by the Institutional Review Board on Human Research of the Faculty of Medicine, Chulalongkorn University with certificate of approval number 814/2020. Written informed consent was obtained from all subjects ≥ 18 years old who were informed of the risks and signed a consent form before enrolling in the study.
Isolation of PBMCs from recovered COVID-19 patients
Blood was collected from 10 hospitalized and recovered COVID-19 patients with high IgG titers against the SARS-CoV-2 spike protein from June to July 2020.
Generation of human antibodies against SARS-CoV-2
To generate human antibodies using hybridoma technology, isolated B cells from recovered COVID-19 patients were transformed by Epstein-Barr virus (EBV) obtained from the B95-8 lymphoblastoid cell line supernatant.
Discussion
In our present study, we successfully generated human monoclonal antibodies using hybridoma technology and successfully isolated the human hybridoma clones producing antibodies against the RBD protein (wild-type) of SARS-CoV-2. We characterized mAbs that were isolated from B cells of recovered COVID-19 patients.
Acknowledgments
We are grateful to the volunteer for participating in the study. We thank the Institute of Biological Products, Department of Medical Sciences, Ministry of Public Health for PRNT results. We also thank Praneet Opanasopit and the team from Department of Pharmaceutical Technology, Faculty of Pharmacy, Silpakorn University for the formulation of the intranasal administration cocktail antibody therapy. We are grateful to the MAX IV laboratory in Lund for providing beamtime at BioMAX. We also want to thank Dr. Ana Gonzales for excellent support during the beamtime.
Citation: Boonkrai C, Cotrone TS, Chaisuriyong W, Tantawichien T, Thisyakorn U, Fernandez S, et al. (2023) Efficacy of the combination of monoclonal antibodies against the SARS-CoV-2 Beta and Delta variants. PLoS ONE 18(5): e0284173. https://doi.org/10.1371/journal.pone.0284173
Editor: Faten Abdelaal Okda, St Jude Children’s Research Hospital, UNITED STATES
Received: October 4, 2022; Accepted: March 25, 2023; Published: May 4, 2023.
Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
Data Availability: All relevant data are within the manuscript and its Supporting Information files.
Funding: This study was supported by Ratchadapiseksompotch Fund, Faculty of Medicine, Chulalongkorn University, grant number RA(P0)001/64, Health Systems Research Institute Fund, grant number 64-112, Education and Research Management Committee, Thai Red Cross Society fund 2021, and Sirivadhanabhakdi foundation.
Competing interests: The authors have declared that no competing interests exist.
