Preparation and identification of monoclonal antibody against RBD protein of porcine ep-idemic diarrhea virus
10.16303/j.cnki.1005-4545.2025.11.03
- VernacularTitle:猪流行性腹泻病毒RBD蛋白单克隆抗体的制备及鉴定
- Author:
Beilei YU
1
;
Yawen ZOU
1
;
Qing HE
1
;
Dantong LI
1
;
Yifan JIANG
1
;
Zhiyong WANG
1
;
Qian YUAN
1
;
Yi YANG
1
;
Naidong WANG
1
Author Information
1. 湖南农业大学动物医学院兽用蛋白质工程疫苗湖南省重点实验室/兽用疫苗逆向创制湖南省工程研究中心,湖南长沙 410128
- Publication Type:Journal Article
- Keywords:
porcine epidemic diarrhea virus;
recombinant baculovirus;
Sf9 cells;
RBD protein;
mono-clonal antibody
- From:
Chinese Journal of Veterinary Science
2025;45(11):2318-2324
- CountryChina
- Language:Chinese
-
Abstract:
The receptor-binding region(RBD)of the spike protein(S)on the surface of porcine epi-demic diarrhea virus(PEDV)is a critical structural domain mediating viral invasion of host cells and serves as a key target for inducing neutralizing antibodies.In order to prepare antibodies that can be used to study the biological function of PEDV RBD and develop novel diagnostic and thera-peutic methods,recombinant RBD protein expressed in Sf9 insect cells was utilized as an immuno-gen to immunize BALB/c mice.Monoclonal antibodies(mAbs)were generated via hybridoma tech-nology,and positive hybridoma clones were screened using indirect ELISA.The reactivity of the mAbs was subsequently characterized.The results of ELISA,Western blot,and indirect immuno-fluorescence assay(IFA)showed that three monoclonal antibodies screened(3E5,4F9 and 5A5)had good reactivity with the virus and RBD protein.Antibody subtype results showed that 3E5 and 4F9 were of IgG1 subtypes and 5A5 was of IgM subtype.Neutralization assay further revealed that 3E5 monoclonal antibody had viral neutralizing activity.In this study,three monoclonal antibodies against PEDV RBD proteins were successfully prepared,providing the basis for the study of the bi-ological function of RBD proteins,PEDV serologic detection and vaccine development.
