Preparation and application of bovine CD4 monoclonal antibodies.
- Author:
Wunjun KONG
1
;
Yueshu ZHU
1
;
Zhengzhong XU
2
;
Chengkun ZHENG
2
;
Xiang CHEN
3
;
Xinan JIAO
2
Author Information
1. Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
2. Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agricultural Product Quality and Safety, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, China.
3. Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agricultural Product Quality and Safety, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, China. *Corresponding author, E-mail: chenxiang@yzu.edu.cn.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal/isolation & purification*;
Cattle;
CD4 Antigens/genetics*;
Hybridomas/immunology*;
Antibody Specificity/immunology*;
Mice;
Mice, Inbred BALB C;
Enzyme-Linked Immunosorbent Assay;
Fluorescent Antibody Technique, Indirect
- From:
Chinese Journal of Cellular and Molecular Immunology
2025;41(5):450-455
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare monoclonal antibodies against bovine CD4 and identify their basic biological characteristics. Methods Recombinant bovine CD4 (rHis-BoCD4 and rGST-BoCD4) was successfully expressed and purified by constructing a prokaryotic plasmid of bovine CD4 gene. The bovine CD4 monoclonal antibody was produced using hybridoma technology. The subtype and potency of the monoclonal antibody were identified and analyzed by ELISA, while specificity was analyzed through indirect immunofluorescence assay (IFA) and Western-blot. Results Four hybridoma cell lines, namely, 1H4, 6A10, 3F9 and 4G10, stably secreting monoclonal antibodies against BoCD4 were successfully obtained. The subclasses of the monoclonal antibodies subclass 6A10 was IgG2b and the rest of the monoclonal antibodies were of IgM type. Western-blot results showed that the four anti-bovine CD4 mAb strains were able to specifically bind to the bovine CD4 protein expressed in vitro. Indirect immunofluorescence assay showed that four monoclonal antibodies were able to specifically recognize the natural bovine CD4 protein. Flow cytometry assay showed that 3F9 was best to recognize bovine natural CD4 molecules. Conclusion Four monoclonal antibody strains with high specificity to natural bovine CD4 protein were successfully prepared, which lays the foundation for the subsequent studies on the function of bovine CD4 and diagnosis and treatment of bovine T-lymphocyte diseases.
