Preparation of peptide mimotope-based diagnostic antigen of Epstein-Barr virus infection
10.3760/cma.j.issn.1003-9279.2018.05.019
- VernacularTitle: 模拟表位型EB病毒感染诊断抗原的制备
- Author:
Qiudong SU
1
;
Minzhuo GUO
2
;
Feng QIU
1
;
Zhiyuan JIA
1
;
Xueting FAN
1
;
Qingling MENG
1
;
Ruiguang TIAN
1
;
Shengli BI
1
;
Yao YI
1
;
Junmei YANG
3
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
2. Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026, China
3. Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou, 450053, China
- Publication Type:Journal Article
- Keywords:
Peptide minotope;
Epstein-Barr virus;
Prokaryotic expression;
Chromatography purification
- From:
Chinese Journal of Experimental and Clinical Virology
2018;32(5):538-542
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare peptide minotope-based recombinant diagnostic antigen of Epstein-Barr virus (EBV) infection and evaluate its antigenicity preliminarily.
Methods:With Trx at the N-terminal and His tag at the C-terminal, the peptide minotope of EBV (GP125, F1, A2, A3C2) was expressed in Escherichia coli and purified by affinity and anion exchange chromatography (designated 'H58’); based on antigenicity of H58 identified by Western blotting (WB), we constructed and evaluated a novel early diagnostic ELISA for EBV infection.
Results:The soluble H58 protein with high concentration (2.8 mg/ml) and purity (99.01) was obtained; WB analysis found that there was an obvious band (28 ×103) on the NC membrane, using H58 anti-Trx monoclonal antibody or acute-phase sera of EBV infection as the first antibody. With the novel ELISA, 50 positive sera of EBV infection and 50 negative sera were detected, displaying that the grouping of OD value of positive serum (95%CI: 1.233-1.489) and negative serum (95%CI: 0.113-0.159) was different (P<0.05) with the sensitivity 98.0%, specificity 96.0% and kappa value 0.940.
Conclusions:By E. coli expression and affinity and ion exchange chromatography purification, the peptide minotope-based recombinant diagnostic antigen of EBV infection was obtained with excellent antigenicity, which could be applied for serological detection of EBV infection.
