Construction and identification of multiple epitope antigens of norovirus based on bioinformatics
10.3969/j.issn.1000-484X.2024.11.025
- VernacularTitle:基于生物信息学诺如病毒多表位抗原的构建及鉴定
- Author:
Xue DU
1
;
Yinzhen ZHAO
;
Yiqing ZHANG
;
Xiaojun WANG
;
Xudong WANG
;
Lanying GUO
;
Yunlong WANG
Author Information
1. 新乡医学院,新乡 453000
- Keywords:
Norovirus;
Bioinformatics analysis;
Multi-epitope antigen
- From:
Chinese Journal of Immunology
2024;40(11):2391-2398
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To design a multi-epitope antigen of norovirus(NoV)based on bioinformatics technology and to pre-pare and characterize it.Methods:Bioinformatics methods were used to construct and analyze the NoV multi-epitope antigen NoV-ZH.Recombinant proteins were prepared and characterized by prokaryotic expression system,and monoclonal antibodies were prepared by animal immunization and hybridoma technology,and initially applied in colloidal gold platform.Results:The designed multi-epitope antigen had a large proportion of random curls in the secondary structure,with theoretical molecular mass and isoelectric point(PI)of 13.1 ku and 7.16,which were stable and hydrophilic.It had good immunogenicity and could activate humoral and cellular immune re-sponses.The proteins prepared by ligating pET-28a(+)and pET-32a vectors with antigenic sequences were expressed as inclusion body proteins and soluble proteins,respectively.A pair of paired antibodies was obtained by animal immunization and hybridoma tech-nique,and applied to colloidal gold test strips with a sensitivity of 0.5 ng/ml,and the test strips could specifically bind two genotypes of NoV recombinant capsid proteins.Conclusion:The successful preparation and characterization of multi-epitope antigen of norovirus provides a reference for the subsequent exploration of NoV universal detection targets and the development of diagnostic raw materials.
