Epitope mapping of Canine distemper virus phosphoprotein by monoclonal antibodies.
The gene for phosphoprotein (P) of CDV encodes three different proteins, P, V, and C. The P protein is involved in viral gene transcription and replication. In the present study, we produced MAbs against a unique domain of the CDV-P protein, from aa 232 to 507, and determined their antigenic sites. By immunizing BALB/c mice with the recombinant P protein-specific fragment, we obtained six MAbs. Competitive binding inhibition assays revealed that they recognized two distinct regions of the P protein. Western blot analysis and immunofluorescence assays using deletion mutants of the unique C-terminus of the CDV-P protein revealed that all MAbs recognized a central short region (aa 233-303) of the CDV-P protein. In addition, linear and conformational epitopes have been determined, and at least four antigenic sites exist in the P protein central region. Furthermore, four of the MAbs were found to react with the P protein of recent Japanese field isolates but not with that of the older CDV strains, including a vaccine strain. Thus, these MAbs could be clinically useful for quick diagnosis during the CDV outbreaks.
Sugai
Akihiro
A
Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan.
Kooriyama
Takanori
T
Sato
Hiroki
H
Yoneda
Misako
M
Kai
Chieko
C
D000818:Animals; D000911:Antibodies, Monoclonal; D019556:COS Cells; D002144:Callithrix; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004217:Distemper Virus, Canine / Q000737:chemistry* / Q000276:immunology*; D000939:Epitopes / Q000032:analysis / Q000737:chemistry*; D005784:Gene Amplification; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D017384:Sequence Deletion; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D000939:Epitopes; D010446:Peptide Fragments; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins