Defucosylation of anti-West Nile virus NS1 antibody enhances ADCC
10.3760/cma.j.cn112309-20241018-00354
- VernacularTitle:去岩藻糖增强抗西尼罗病毒NS1抗体的ADCC效应
- Author:
Wanlu ZHU
1
;
Lingli WU
;
Nan CHEN
;
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:
CRISPR/Cas9 system;
Gene knockout;
Anti-fucose antibody;
ADCC;
SLC35C1
- From:
Chinese Journal of Microbiology and Immunology
2025;45(9):740-745
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To obtain fucose-free anti-West Nile virus nonstructural protein 1 (NS1) antibody and evaluate its antibody-dependent cell-mediated cytotoxicity (ADCC).Methods:The guanosine diphosphate-fucose transporter SLC35C1 in CHO cells was knocked out using CRISPR/Cas9 gene editing technology to obtain the fucose-free cell line CHO SLC35C1 -/-. CHO SLC35C1 -/- cells were used to produce fucose-free anti-West Nile virus NS1 antibodies. The binding abilities of the antibodies to the target antigen of West Nile virus NS1 protein and the human high-affinity IgG Fc receptor hFcγRⅠ (hCD64) were detected by ELISA and flow cytometry, respectively. The ADCC activity of the antibodies was detected by ADCC reporter gene assay. One-way analysis of variance was used for statistical analysis. Results:CHO SLC35C1 -/- cells expressed green fluorescent protein but not Lens culinaris agglutinin. The anti-West Nile virus NS1 antibodies produced by CHO SLC35C1 -/- cells with a fucose content of 0.22% could bind to West Nile virus NS1 protein in a concentration-dependent manner. Compared with the wild-type antibodies, the fucose-free anti-West Nile virus NS1 antibodies showed a stronger binding ability to hFcγRⅠ(hCD64), as indicated by a significant increase in fluorescence intensity. The ADCC reporter gene assay showed that the fucose-free anti-West Nile virus NS1 antibodies had increased activity as compared with the wild-type antibodies ( P<0.001). Conclusion:The fucose-free anti-West Nile virus NS1 antibodies may be used to protect against West Nile virus infection.
