Characterization and application of monoclonal antibodies to bovine viral diarrhea virus nonstructural protein 5A.
Bovine viral diarrhea virus (BVDV) nonstructural protein 5A (NS5A) is one the least studied of the BVDV proteins. Therefore, to develop a tool for unraveling the functions performed by BVDV NS5A, monoclonal antibodies (MAbs) were generated by fusion of myeloma cells with spleen cells from mice immunized with recombinant E. coli-expressed GST-NS5A protein. Two MAbs (1H12 and 2F9) were established on the basis of immunofluorescence and Western blot analysis. Both the MAbs were of IgG1 subclass and recognized an epitope clustered within the N-terminal region of NS5A. Furthermore, the MAb 1H12 was used successfully to detect NS5A protein in BVDV field isolates belonging to genotypes 1 and 2. Temporal expression pattern studies during an infectious cycle revealed that BVDV NS5A could be detected 12-60 h postinfection. Confocal microscopy studies showed a cytoplasmic staining pattern and revealed that NS5A is localized on the endoplasmic reticulum membrane in BVDV infected cells.
Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Muhammad Atif
MA
Zahoor
Daisuke
D
Yamane
Yassir Mahgoub
YM
Mohamed
Kyousuke
K
Kobayashi
Kentaro
K
Kato
Yukinobu
Y
Tohya
Hiroomi
H
Akashi
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000378:metabolism*; D002417:Cattle; D002459:Cell Fusion; D002460:Cell Line; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D005838:Genotype; D051379:Mice; D008807:Mice, Inbred BALB C; D013997:Time Factors; D017361:Viral Nonstructural Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D017361:Viral Nonstructural Proteins