Expression and purification of different segments from HCoV-NL63 nucleocapsid protein and their application in detection of antibodies.
- Author:
Min ZHAO
1
;
Ting-Ying ZHANG
;
Wei-Min ZHOU
;
Guo-Xia ZHAO
;
Ling-Lin ZHANG
;
Ji-Min GAO
;
Wen-Jie TAN
Author Information
1. Institute of Medical Virology, Wenzhou Medical College, Wenzhou 325000, China.
- Publication Type:Journal Article
- MeSH:
Adult;
Antibodies, Viral;
blood;
Blotting, Western;
Coronavirus;
chemistry;
immunology;
Humans;
Nucleocapsid Proteins;
genetics;
immunology;
isolation & purification;
Peptide Fragments;
genetics;
isolation & purification;
Recombinant Proteins;
biosynthesis;
immunology;
isolation & purification;
Serologic Tests
- From:
Chinese Journal of Virology
2011;27(3):244-249
- CountryChina
- Language:Chinese
-
Abstract:
Prokaryotic expression plasmids carrying N-terminal(1-163aa) and C-terminal(141-306aa) gene of HCoV-NL63 nucleocapsid protein were constructed with pET-30a(+) vector. Consequently, we prepared two purified proteins, Np and Cp, respectively, and established a Western blotting-based line assay (WBLA) for detection of antibodies against HCoV-NL63 using three purified proteins: Np , Cp and Nf, a full-length HCoV-NL63 nucleocapsid protein as previously reported. We detected anti-HCoV-NL63 antibodies among 50 sera samples collected from adult for health-examination by WBLA. The results showed that: 25 (50%), 27 (54%), 36 (72%) of 50 sera were indentified as anti-HCoV-NL63 antibody positive when the antigen was from Nf, Np and Cp, respectively. Among these sera with positive anti-HCoV-NL63 antibody,Cp showed highest antibody positive rate in WBLA,and consistent rates of detection were 64% between Nf and Np, 54% between Nf and Cp, 54% between Np and Cp. Our study provides the foundation for development of HCoV-NL63 serological detection reagents and an experimental tool for immunological research of HCoV-NL63 infection.
