Preparation of monoclonal antibody against PRRSV-2 N protein and identification of antigenic epitopes
10.16303/j.cnki.1005-4545.2025.01.03
- VernacularTitle:2型PRRSV N蛋白单克隆抗体的制备及其抗原表位的鉴定
- Author:
Yanli PANG
1
;
Jianguang QIN
1
;
Muyang LIU
1
;
Tongwei REN
1
;
Jiaqi LIU
1
;
Lingshan ZHOU
1
;
Ying CHEN
1
;
Kang OUYANG
1
;
Weijian HUANG
1
;
Zuzhang WEI
1
Author Information
1. 广西大学动物科学技术学院广西高校动物疫病预防与控制重点实验室,广西南宁 530005
- Publication Type:Journal Article
- Keywords:
PRRSV;
nucleocapsid protein;
monoclonal antibody;
antigenic epitope
- From:
Chinese Journal of Veterinary Science
2025;45(1):16-21,45
- CountryChina
- Language:Chinese
-
Abstract:
To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.
