Open AccessCharacterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
Author(s) -
Xin Zhang,
Xin Zhao,
Hui Dong,
Yali Zhu,
Hongyan Shi,
Jianfei Chen,
Da Shi,
Li Feng
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0163920
Subject(s) - monoclonal antibody , immunoprecipitation , virology , epitope , coronavirus , biology , immunofluorescence , epitope mapping , linker , microbiology and biotechnology , antibody , blot , cytoplasm , antigen , antigenicity , viral structural protein , conformational epitope , viral replication , virus , gene , covid-19 , viral entry , biochemistry , genetics , medicine , disease , pathology , computer science , infectious disease (medical specialty) , operating system
The transmissible gastroenteritis virus (TGEV) nucleocapsid (N) protein plays important roles in the replication and translation of viral RNA. The present study provides the first description of two monoclonal antibodies (mAbs) (5E8 and 3D7) directed against the TGEV N protein linker region (LKR) and carboxyl terminal domain (CTD). The mAbs 5E8 and 3D7 reacted with native N protein in western blotting and immunofluorescence assay (IFA). Two linear epitopes, 189 SVEQAVLAALKKLG 202 and 246 VTRFYGARSSSA 257 , located in the LKR and CTD of TGEV N protein, respectively, were identified after truncating the protein and applying a peptide scanning technique. Using mAb 5E8, we observed that the N protein was expressed in the cytoplasm during TGEV replication and that the protein could be immunoprecipitated from TGEV-infected PK-15 cells. The mAb 5E8 can be applied for different approaches to diagnosis of TGEV infection. In addition, the antibodies represent useful tools for investigating the antigenic properties of the N protein.