Preparation and application of CD318 monoclonal antibody.
- Author:
Ke CHAO
1
;
Ziyang WANG
2
;
Jie ZHAO
3
;
Meijia YANG
4
,
5
Author Information
1. National Engineering Laboratory for Internet Medical Systems and Applications, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
2. Outpatient Department, the Hospital of Henan Provincial Corps of the Chinese people's Armed Police Force, Zhengzhou 450052, China.
3. National Engineering Laboratory for Internet Medical Systems and Applications, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Corresponding authors, E-mail: zhaojie@zzu.edu.cn.
4. National Engineering Laboratory for Internet Medical Systems and Applications, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. *Corresponding authors, E-mail: meijiayang2293@
5. com.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal/biosynthesis*;
Mice;
Antibodies, Bispecific/immunology*;
Humans;
Mice, Inbred BALB C;
Antibody Specificity/immunology*;
CD3 Complex/immunology*;
Antigens, CD/genetics*;
T-Lymphocytes/immunology*;
Hybridomas/immunology*;
Female
- From:
Chinese Journal of Cellular and Molecular Immunology
2025;41(9):818-826
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare CD318-specific monoclonal antibodies and evaluate their specificity, affinity, and application in immunological detection, laying the foundation for the development of CD318-targeted antibody drugs. MethodsCD318 protein was expressed and purified, and was used as an antigen to immunize mice, then mice with higher antiserum titers were screened. We prepared CD318-specific monoclonal antibodies through cell fusion and monoclonal screening, and the specificity, affinity, and application of the obtained monoclonal antibodies in immunological assays were evaluated. Then we constructed a CD318/CD3-targeting bispecific antibody and assessed its impact on T-cell cytotoxicity. Results Thirteen monoclonal antibodies were successfully generated, with the hybridoma clone 13-8-G2 exhibiting the highest titer, strongest specificity, and broadest applicability. The antibody was identified as an IgG1 isotype with a kappa light chain. The variable region of the light chain measured 318 bp, while the heavy chain variable region was 357 bp, yielding an affinity constant of approximately 7.68×109. The specificity of CD318 was confirmed using flow cytometry and immunofluorescence assays. Additionally, a CD318/CD3-targeting bispecific antibody was constructed using the variable regions of this CD318 monoclonal antibody, which demonstrated enhanced T-cell cytotoxicity. Conclusion High-affinity and highly specific CD318 monoclonal antibodies were successfully prepared, laying a foundation for the development of therapeutic antibodies targeting CD318.
