Preparation and identification of monoclonal antibodies against S1 protein of por-cine epidemic diarrhea virus
10.16303/j.cnki.1005-4545.2025.04.01
- VernacularTitle:猪流行性腹泻病毒S1蛋白单克隆抗体的制备与鉴定
- Author:
Chaofan LIU
1
;
Jing REN
;
Feiyan WANG
;
Xiaojing SHI
;
Jialu HOU
;
Yuwei ZHAI
;
Chen YUAN
;
Qinye SONG
Author Information
1. 河北农业大学动物医学院河北省兽医生物技术创新中心,河北保定 071000
- Publication Type:Journal Article
- Keywords:
porcine epidemic diarrhea virus(PEDV);
S1 protein;
monoclonal antibodies
- From:
Chinese Journal of Veterinary Science
2025;45(4):627-633
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to prepare monoclonal antibody to S1 protein of porcine epidemic diar-rhea virus(PEDV).E.coli expression system and affinity chromatography were used to success-fully obtain purified recombinant PEDV S1 protein.After immunizing BALB/c mice,hybridoma technology and indirect ELISA were used to prepare and screen positive hybridoma cells.Finally,ascites antibodies were prepared by in vivo induction method.ELISA results showed that a total of 4 hybridoma cell lines with anti-PEDV S1 monoclonal antibody were screened,and they were named E6,G3,H6 and F2.The supernatant titers of all 4 hybridoma cell lines reached 1∶6 400.The monoclonal antibody H6 with higher antibody titers and more stable antibody secretion was selected for antibody type identification.It was found that monoclonal antibody H6 belongs to the IgG1 subclass and the light chain is the λ chain.The antibody titers that induced mouse ascites were 1∶106 and without cross-reaction with other proteins.Western blot results showed that the monoclonal antibody exhibited specific bands at 38 kDa with the recombinant S1 protein,PEDV QY2016,and PEDV CV777 strains.The IFA results also showed that the monoclonal antibody reacted with cells infected with PEDV QY2016 and PEDV CV777 strains,exhibiting a green fluo-rescent signal.The affinity constant of monoclonal antibody H6 was K=1.75×107 moL/L,indica-ting that the H6 strain had a good affinity and could be used for the development of subsequent di-agnostic antibodies.In summary,this study successfully prepared monoclonal antibodies that can specifically recognize PEDV S1 protein,which can be used for the antigen detection of PEDV and providing important test materials for the research of PEDV detection methods.
