title: Confirmation of a new conserved linear epitope of Lyssavirus nucleoprotein.
abstract: Bioinformatics analysis was used to predict potential epitopes of Lyssavirus nucleoprotein and highlighted some distinct differences in the quantity and localization of the epitopes disclosed by epitope analysis of monoclonal antibodies against Lyssavirus nucleoprotein. Bioinformatics analysis showed that the domain containing residues 152-164 of Lyssavirus nucleoprotein was a conserved linear epitope that had not been reported previously. Immunization of two rabbits with the corresponding synthetic peptide conjugated to the Keyhole Limpe hemocyanin (KLH) macromolecule resulted in a titer of anti-peptide antibody above 1:200,000 in rabbit sera as detected by indirect enzyme-linked immunosorbent assay (ELISA). Western blot analysis demonstrated that the anti-peptide antibody recognized denatured Lyssavirus nucleoprotein in sodium dodecylsulfonate-polyacrylate gel electrophoresis (SDS-PAGE). Affinity chromatography purification and FITC-labeling of the anti-peptide antibody in rabbit sera was performed. FITC-labeled anti-peptide antibody could recognize Lyssavirus nucleoprotein in BSR cells and canine brain tissues even at a 1:200 dilution. Residues 152-164 of Lyssavirus nucleoprotein were verified as a conserved linear epitope in Lyssavirus.
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, Department of Viral Encephalitis, Institute for Viral Disease Control and Prevention, Chinese Centers for Disease Control and Prevention, No. 155 Changbai Road, Changping District, Beijing 102206, China.
authors.forename: Lv
authors.initials: L
authors.lastname: Xinjun
authors.forename: Ma
authors.initials: M
authors.lastname: Xuejun
authors.forename: Wang
authors.initials: W
authors.lastname: Lihua
authors.forename: Li
authors.initials: L
authors.lastname: Hao
authors.forename: Shen
authors.initials: S
authors.lastname: Xinxin
authors.forename: Yu
authors.initials: Y
authors.lastname: Pengcheng
authors.forename: Tang
authors.initials: T
authors.lastname: Qing
authors.forename: Liang
authors.initials: L
authors.lastname: Guodong
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western; D001921:Brain / Q000821:virology; D019295:Computational Biology / Q000379:methods; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D007114:Immunization / Q000379:methods; D018114:Lyssavirus / Q000235:genetics / Q000276:immunology*; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; D009698:Nucleoproteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic