title: Monoclonal antibodies against the nucleocapsid proteins of henipaviruses: production, epitope mapping and application in immunohistochemistry.
abstract: Four monoclonal antibodies (mAbs) were generated by immunizing BALB/C mice with recombinant nucleocapsid protein (N) of Nipah virus (NiV) and Hendra virus (HeV) expressed in E. coli. Two mAbs each were obtained for the HeV N and NiV N, respectively. All four mAbs displayed specific reactivity with the recombinant N proteins of both viruses by western blot, which was further confirmed by immunofluorescent antibody assay using fixed insect cells infected with recombinant baculoviruses expressing either the HeV or NiV N protein. Epitope mapping using a 12-mer random peptide phage display library revealed two linear antigenic sites of the henipavirus N proteins, KLxR (aa 17-20) and FKREM (aa 446-450), which have not been reported previously. Two of the mAbs were able to specifically recognize HeV antigens by immunohistochemical staining of lung tissue sections of a horse experimentally infected with HeV. These reagents will be a useful addition to the collection of tools essential for further research and improvement in diagnosis of henipaviruses.
authors.affiliation: Institute of Veterinary Sciences, Academy of Military Medical Sciences, Changchun, China.
authors.forename: C
authors.initials: C
authors.lastname: Xiao
authors.forename: Y
authors.initials: Y
authors.lastname: Liu
authors.forename: Y
authors.initials: Y
authors.lastname: Jiang
authors.forename: D E
authors.initials: DE
authors.lastname: Magoffin
authors.forename: H
authors.initials: H
authors.lastname: Guo
authors.forename: H
authors.initials: H
authors.lastname: Xuan
authors.forename: G
authors.initials: G
authors.lastname: Wang
authors.forename: L-F
authors.initials: LF
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Tu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D018604:Epitope Mapping; D005260:Female; D045404:Hendra Virus / Q000276:immunology*; D045464:Henipavirus Infections / Q000473:pathology / Q000821:virology; D006734:Horse Diseases / Q000473:pathology / Q000821:virology; D006736:Horses; D007150:Immunohistochemistry / Q000379:methods*; D008168:Lung / Q000473:pathology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D045405:Nipah Virus / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000276:immunology; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D011994:Recombinant Proteins