Monoclonal antibodies against the nucleocapsid proteins of henipaviruses: production, epitope mapping and application in immunohistochemistry.
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.
Xiao
C
C
Institute of Veterinary Sciences, Academy of Military Medical Sciences, Changchun, China.
Liu
Y
Y
Jiang
Y
Y
Magoffin
D E
DE
Guo
H
H
Xuan
H
H
Wang
G
G
Wang
L-F
LF
Tu
C
C
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D011994:Recombinant Proteins