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Isolation of serotype-specific antibodies against Dengue virus non-structural protein 1 using phage display and application in a multiplexed serotyping assay

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dc.contributor.author Lebani, Kebaneilwe
dc.contributor.author Jones, Martina L
dc.contributor.author Watterson, Daniel
dc.contributor.author Ranzoni, Andrea
dc.contributor.author Traves, Renee J
dc.contributor.author Young, Paul R
dc.contributor.author Mahler, Stephen M
dc.date.accessioned 2021-02-19T10:40:01Z
dc.date.available 2021-02-19T10:40:01Z
dc.date.issued 2017-07-06
dc.identifier.citation Lebani,K. et al (2017) Isolation of serotype-specific antibodies against dengue virus non-structural protein 1 using phage display and application in a multiplexed serotyping assay. PLoS ONE 12(7), e0180669. https://doi.org/10.1371/journal.pone.0180669 en_US
dc.identifier.uri http://repository.biust.ac.bw/handle/123456789/272
dc.description.abstract The multidimensional nature of dengue virus (DENV) infections, which can be caused by four distinct serotypes of the virus, complicates the sensitivity of assays designed for the diagnosis of infection. Different viral markers can be optimally detected at different stages of infection. Of particular clinical importance is the early identification of infection, which is pivotal for disease management and the development of blood screening assays. Non-structural protein 1 (NS1) is an early surrogate marker of infection and its detection in serum coincides with detectable viraemia. The aim of this work was to isolate and characterise serotype-specific monoclonal antibodies that bind to NS1 for each of the four DENV serotypes. This was achieved using phage display and a subtractive biopanning strategy to direct the antibody selection towards serotype-specific epitopes. This antibody isolation strategy has advantages over immunisation techniques where it is difficult to avoid antibody responses to cross-reactive, immunodominant epitopes. Serotype specificity to recombinant antigen for each of the antibodies was confirmed by Enzyme Linked Immunosorbent Assay (ELISA) and Surface Plasmon Resonance. Confirmation of binding to native DENV NS1 was achieved using ELISA and immunofluorescence assay on DENV infected Vero cells. No cross-reactivity with Zika or Kunjin viruses was observed. A previously isolated pan-reactive antibody that binds to an immunodominant epitope was able to pair with each of the serotype-specific antibodies in a sandwich ELISA, indicating that the serotype specific antibodies bind to epitopes which are all spatially distinct from the immunodominant epitope. These antibodies were suitable for use in a multiplexed assay for simultaneous detection and serotyping of DENV NS1 in human serum. This work demonstrates that phage display coupled with novel biopanning strategies is a valuable in vitro methodology for isolation of binders that can discern amongst antigens with high homology for diagnostic applicability. en_US
dc.language.iso en en_US
dc.publisher PLOS ONE en_US
dc.subject Viral structural proteins en_US
dc.subject Bacteriophages en_US
dc.title Isolation of serotype-specific antibodies against Dengue virus non-structural protein 1 using phage display and application in a multiplexed serotyping assay en_US
dc.description.level phd en_US
dc.description.accessibility unrestricted en_US
dc.description.department bsb en_US


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