High affinity human antibody fragments to dengue virus non-structural protein 3.
BACKGROUND: The enzyme activities catalysed by flavivirus non-structural protein 3 (NS3) are essential for virus replication. They are distributed between the N-terminal protease domain in the first one-third and the C-terminal ATPase/helicase and nucleoside 5' triphosphatase domain which forms...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-01-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC2976680?pdf=render |
| _version_ | 1811279336566685696 |
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| author | Nicole J Moreland Moon Y F Tay Elfin Lim Prasad N Paradkar Danny N P Doan Yin Hoe Yau Susana Geifman Shochat Subhash G Vasudevan |
| author_facet | Nicole J Moreland Moon Y F Tay Elfin Lim Prasad N Paradkar Danny N P Doan Yin Hoe Yau Susana Geifman Shochat Subhash G Vasudevan |
| author_sort | Nicole J Moreland |
| collection | DOAJ |
| description | BACKGROUND: The enzyme activities catalysed by flavivirus non-structural protein 3 (NS3) are essential for virus replication. They are distributed between the N-terminal protease domain in the first one-third and the C-terminal ATPase/helicase and nucleoside 5' triphosphatase domain which forms the remainder of the 618-aa long protein. METHODOLOGY/PRINCIPAL FINDINGS: In this study, dengue full-length NS3 protein with residues 49 to 66 of NS2B covalently attached via a flexible linker, was used as bait in biopanning with a naïve human Fab phage-display library. Using a range of truncated constructs spanning the NS2B cofactor region and the full-length NS3, 10 unique Fab were identified and characterized. Of these, monoclonal Fab 3F8 was shown to bind α3″ (residues 526 through 531) within subdomain III of the helicase domain. The antibody inhibits the ATPase and helicase activites of NS3 in biochemical assays and reduces DENV replication in HEK293 cells that were previously transfected with Fab 3F8 compared with mock transfected cells. CONCLUSIONS/SIGNIFICANCE: Antibodies such as 3F8 are valuable tools for studying the molecular mechanisms of flaviviral replication and for the monospecific detection of replicating dengue virus in vivo. |
| first_indexed | 2024-04-13T00:52:49Z |
| format | Article |
| id | doaj.art-051a3d034d624126913cd52bb6d25081 |
| institution | Directory Open Access Journal |
| issn | 1935-2735 |
| language | English |
| last_indexed | 2024-04-13T00:52:49Z |
| publishDate | 2010-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-051a3d034d624126913cd52bb6d250812022-12-22T03:09:48ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27352010-01-01411e88110.1371/journal.pntd.0000881High affinity human antibody fragments to dengue virus non-structural protein 3.Nicole J MorelandMoon Y F TayElfin LimPrasad N ParadkarDanny N P DoanYin Hoe YauSusana Geifman ShochatSubhash G VasudevanBACKGROUND: The enzyme activities catalysed by flavivirus non-structural protein 3 (NS3) are essential for virus replication. They are distributed between the N-terminal protease domain in the first one-third and the C-terminal ATPase/helicase and nucleoside 5' triphosphatase domain which forms the remainder of the 618-aa long protein. METHODOLOGY/PRINCIPAL FINDINGS: In this study, dengue full-length NS3 protein with residues 49 to 66 of NS2B covalently attached via a flexible linker, was used as bait in biopanning with a naïve human Fab phage-display library. Using a range of truncated constructs spanning the NS2B cofactor region and the full-length NS3, 10 unique Fab were identified and characterized. Of these, monoclonal Fab 3F8 was shown to bind α3″ (residues 526 through 531) within subdomain III of the helicase domain. The antibody inhibits the ATPase and helicase activites of NS3 in biochemical assays and reduces DENV replication in HEK293 cells that were previously transfected with Fab 3F8 compared with mock transfected cells. CONCLUSIONS/SIGNIFICANCE: Antibodies such as 3F8 are valuable tools for studying the molecular mechanisms of flaviviral replication and for the monospecific detection of replicating dengue virus in vivo.http://europepmc.org/articles/PMC2976680?pdf=render |
| spellingShingle | Nicole J Moreland Moon Y F Tay Elfin Lim Prasad N Paradkar Danny N P Doan Yin Hoe Yau Susana Geifman Shochat Subhash G Vasudevan High affinity human antibody fragments to dengue virus non-structural protein 3. PLoS Neglected Tropical Diseases |
| title | High affinity human antibody fragments to dengue virus non-structural protein 3. |
| title_full | High affinity human antibody fragments to dengue virus non-structural protein 3. |
| title_fullStr | High affinity human antibody fragments to dengue virus non-structural protein 3. |
| title_full_unstemmed | High affinity human antibody fragments to dengue virus non-structural protein 3. |
| title_short | High affinity human antibody fragments to dengue virus non-structural protein 3. |
| title_sort | high affinity human antibody fragments to dengue virus non structural protein 3 |
| url | http://europepmc.org/articles/PMC2976680?pdf=render |
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