Preparation of monoclonal antibodies against 3D protein of EV71 based on HBc particles as expression vector
10.3969/j.issn.1000-484X.2017.09.013
- VernacularTitle:以乙肝病毒核心蛋白为载体制备肠道病毒71型非结构蛋白3D单克隆抗体
- Author:
Yongchao LI
;
Rui ZHU
;
Longfa XU
;
Yangtao WU
;
Huan ZHAO
;
Kun WU
;
Dongxiao LIU
;
Tong CHENG
;
Ningshao XIA
- Keywords:
Hepatitis B virus core protein;
Enterovirus 71;
Non-structural protein 3D;
Monoclonal antibody
- From:
Chinese Journal of Immunology
2017;33(9):1341-1345
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.
