TY - JOUR
T1 - Generation and characterization of mouse monoclonal antibodies against NS4B protein of dengue virus
AU - Xie, Xuping
AU - Zou, Jing
AU - Wang, Qing Yin
AU - Noble, Christian G.
AU - Lescar, Julien
AU - Shi, Pei Yong
N1 - Funding Information:
We thank Andy Yip and other colleagues for help during the course of this study. This project was partially supported by the TCR flagship “STOP Dengue” Program from National Medical Research Council in Singapore.
PY - 2014/2
Y1 - 2014/2
N2 - Dengue virus (DENV) non-structural protein 4B (NS4B) has been demonstrated to be an attractive antiviral target. Due to its nature as an integral membrane protein, NS4B remains poorly characterized. In this study, we generated and characterized two monoclonal antibodies (mAb) that selectively bind to DENV NS4B protein. One mAb, 10-3-7, is specific for DENV-2 NS4B, and its epitope was mapped to residues 5-15 of NS4B. The other mAb, 44-4-7, cross-reacts with all the four serotypes of DENV NS4B, and its epitope was mapped to residues 141-147 of NS4B. Using the mAbs, we probed the intracellular orientation of the epitopes of NS4B by an epitope accessibility assay. The results showed that the N-terminus of NS4B is located in the ER lumen, whereas amino acids 130-148 of NS4B are located in the cytosol. The study demonstrates that the two anti-NS4B mAbs will be useful for future structural and functional analyses of DENV NS4B.
AB - Dengue virus (DENV) non-structural protein 4B (NS4B) has been demonstrated to be an attractive antiviral target. Due to its nature as an integral membrane protein, NS4B remains poorly characterized. In this study, we generated and characterized two monoclonal antibodies (mAb) that selectively bind to DENV NS4B protein. One mAb, 10-3-7, is specific for DENV-2 NS4B, and its epitope was mapped to residues 5-15 of NS4B. The other mAb, 44-4-7, cross-reacts with all the four serotypes of DENV NS4B, and its epitope was mapped to residues 141-147 of NS4B. Using the mAbs, we probed the intracellular orientation of the epitopes of NS4B by an epitope accessibility assay. The results showed that the N-terminus of NS4B is located in the ER lumen, whereas amino acids 130-148 of NS4B are located in the cytosol. The study demonstrates that the two anti-NS4B mAbs will be useful for future structural and functional analyses of DENV NS4B.
KW - Dengue virus NS4B protein
KW - Flavivirus replication
KW - Membrane protein
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U2 - 10.1016/j.virol.2013.12.025
DO - 10.1016/j.virol.2013.12.025
M3 - Article
C2 - 24503088
AN - SCOPUS:84891897588
SN - 0042-6822
VL - 450-451
SP - 250
EP - 257
JO - Virology
JF - Virology
ER -