title: [Expression and purification of different segments from HCoV-NL63 nucleocapsid protein and their application in detection of antibodies].
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.
authors.affiliation: Institute of Medical Virology, Wenzhou Medical College, Wenzhou 325000, China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhao
authors.forename: Ting-Ying
authors.initials: TY
authors.lastname: Zhang
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Zhou
authors.forename: Guo-Xia
authors.initials: GX
authors.lastname: Zhao
authors.forename: Ling-Lin
authors.initials: LL
authors.lastname: Zhang
authors.forename: Ji-Min
authors.initials: JM
authors.lastname: Gao
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Tan
mesh_terms: D000328:Adult; D000914:Antibodies, Viral / Q000097:blood*; D015153:Blotting, Western; D017934:Coronavirus / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D010446:Peptide Fragments / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D012698:Serologic Tests
chemical_list: D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins