TY - JOUR
T1 - Characterization and application of monoclonal antibodies to bovine viral diarrhea virus nonstructural protein 5A
AU - Zahoor, Muhammad Atif
AU - Yamane, Daisuke
AU - Mohamed, Yassir Mahgoub
AU - Kobayashi, Kyousuke
AU - Kato, Kentaro
AU - Tohya, Yukinobu
AU - Akashi, Hiroomi
PY - 2009/10/1
Y1 - 2009/10/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=70350651276&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70350651276&partnerID=8YFLogxK
U2 - 10.1007/s00705-009-0505-7
DO - 10.1007/s00705-009-0505-7
M3 - Article
C2 - 19795184
AN - SCOPUS:70350651276
VL - 154
SP - 1745
EP - 1754
JO - Archives of Virology
JF - Archives of Virology
SN - 0304-8608
IS - 11
ER -