Multimerization through PEGylation improves properties of a single-chain variable fragment against West Nile virus
10.3760/cma.j.cn112309-20250514-00150
- VernacularTitle:聚乙二醇化可改善抗西尼罗病毒单链抗体活性
- Author:
Wanlu ZHU
1
;
Lingli WU
;
Huihui JIA
;
Beifen SHEN
;
Jiannan FENG
;
Jun ZHANG
;
He XIAO
Author Information
1. 军事科学院军事医学研究院，北京 100039
- Publication Type:Journal Article
- Keywords:
West Nile virus;
PEGylation;
Single-chain variable fragment;
Multimerization
- From:
Chinese Journal of Microbiology and Immunology
2025;45(11):914-919
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To obtain a polyvalent single-chain variable fragment（scFv）against West Nile virus through PEGylation in order to improve its antigen-binding ability and neutralizing activity.Methods:A scFv carrying a C-terminal cysteine residue（scFvC）was constructed by introducing Cys into the C-terminal of scFv against West Nile virus. Then the multimerization of scFvC was achieved by targeting the thiol group of Cys with maleimide-activated polyethylene glycol. ELISA was used to detect the antigen-binding activity of the multivalent scFvC. Pseudovirus-based neutralization assay was used to evaluate the neutralizing activity of the multivalent scFvC in vitro. One-way analysis of variance was used for statistical analysis. Results:The PEGylated scFvC multimers showed higher antigen-binding ability than the monomeric scFvC. In the pseudovirus-based neutralization assay，both monomeric scFvC and PEGylated scFvC multimers showed good neutralizing activity compared with the control group（ P<0.000 1）. Moreover，the PEGylated scFvC multimers showed a more effective ability to block the pseudovirus infection in target cells（ P<0.05），suggesting that the PEGylated scFvC multimers could enhance their function in vitro through avidity effect. Conclusion:In this study，a scFvC targeting West Nile virus is successfully constructed and its polyvalent form is generated through PEGylation，which improves the antigen-binding and neutralizing activity of the parental scFv.
