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...
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ftdoajarticles:oai:doaj.org/article:051a3d034d624126913cd52bb6d25081 2023-05-15T15:06:58+02:00 High affinity human antibody fragments to dengue virus non-structural protein 3. 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 2010-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0000881 https://doaj.org/article/051a3d034d624126913cd52bb6d25081 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2976680?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0000881 https://doaj.org/article/051a3d034d624126913cd52bb6d25081 PLoS Neglected Tropical Diseases, Vol 4, Iss 11, p e881 (2010) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2010 ftdoajarticles https://doi.org/10.1371/journal.pntd.0000881 2022-12-30T22:10:48Z 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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 4 11 e881 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 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. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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. |
format |
Article in Journal/Newspaper |
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 |
title |
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_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_sort |
high affinity human antibody fragments to dengue virus non-structural protein 3. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pntd.0000881 https://doaj.org/article/051a3d034d624126913cd52bb6d25081 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 4, Iss 11, p e881 (2010) |
op_relation |
http://europepmc.org/articles/PMC2976680?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0000881 https://doaj.org/article/051a3d034d624126913cd52bb6d25081 |
op_doi |
https://doi.org/10.1371/journal.pntd.0000881 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
4 |
container_issue |
11 |
container_start_page |
e881 |
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1766338545556914176 |