Construction of a human anti-SARS-CoV-2 scFv library and identification of broad-spectrum neutralizing antibodies.
- Author:
Huimin YIN
1
;
Hai LYU
1
;
Ying CHI
2
;
Jingxian LIU
2
;
Yongjun JIAO
3
,
4
;
Pingmin WEI
4
,
5
Author Information
1. Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, China.
2. Key Laboratory of Enteric Pathogenic Microorganisms, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China.
3. Key Laboratory of Enteric Pathogenic Microorganisms, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China. *Corresponding authors, E-mail: yongjunjiao@
4. com.
5. Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, China. *Corresponding authors, E-mail: mpw1963@
- Publication Type:Journal Article
- MeSH:
Humans;
Single-Chain Antibodies/genetics*;
SARS-CoV-2/immunology*;
COVID-19/immunology*;
Immunoglobulin G/genetics*;
Antibodies, Viral/genetics*;
Peptide Library;
Spike Glycoprotein, Coronavirus/immunology*;
Antibodies, Neutralizing/immunology*;
Leukocytes, Mononuclear/immunology*;
Broadly Neutralizing Antibodies/immunology*
- From:
Chinese Journal of Cellular and Molecular Immunology
2025;41(2):154-160
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a library of human-derived anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) single-chain variable fragments (scFv) and screen for broad-spectrum neutralizing antibodies to identify candidate molecules for the development of diagnostic and therapeutic agents. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of patients who had recovered from novel coronavirus infection. Total RNA was extracted from these PBMCs and reverse transcribed into cDNA, which was used as a template for constructing a human anti-SARS-CoV-2 scFv library. Phage display technology was used to screen for scFv antibodies specific to the SARS-CoV-2 S protein. Full-length IgG antibodies were synthesized through sequence analysis and human IgG expression, and their binding capacity and neutralizing activity against SARS-CoV-2 were evaluated. Results A human-derived scFv antibody library against SARS-CoV-2 with a capacity of 1.56×107 CFU was successfully constructed. Two specific scFv antibodies were screened from this library and expressed as full-length IgG antibodies (IgG-A10 and IgG-G6). IgG-A10 exhibited strong neutralizing activity against both the original SARS-CoV-2 strain (WT) and the XBB subvariant of the Omicron variant. However, the neutralizing activity of this antibody against the JN.1 sub lineage of the Omicron BA.2.86 variant was moderate. Conclusion This study has successfully constructed a human anti-SARS-CoV-2 scFv antibody library from the peripheral blood of recovered patients, and screened and expressed anti-SARS-CoV-2 IgG antibodies with neutralizing activity, laying a foundation for the prevention, diagnosis, and treatment of SARS-CoV-2 infection.
