Please use this identifier to cite or link to this item: https://doi.org/10.3390/ijms13032618
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dc.titlePhage display approaches for the isolation of monoclonal antibodies against dengue virus envelope domain III from human and mouse derived libraries
dc.contributor.authorMoreland, N.J.
dc.contributor.authorSusanto, P.
dc.contributor.authorLim, E.
dc.contributor.authorTay, M.Y.F.
dc.contributor.authorRajamanonmani, R.
dc.contributor.authorHanson, B.J.
dc.contributor.authorVasudevan, S.G.
dc.date.accessioned2014-11-26T08:29:49Z
dc.date.available2014-11-26T08:29:49Z
dc.date.issued2012-03
dc.identifier.citationMoreland, N.J., Susanto, P., Lim, E., Tay, M.Y.F., Rajamanonmani, R., Hanson, B.J., Vasudevan, S.G. (2012-03). Phage display approaches for the isolation of monoclonal antibodies against dengue virus envelope domain III from human and mouse derived libraries. International Journal of Molecular Sciences 13 (3) : 2618-2635. ScholarBank@NUS Repository. https://doi.org/10.3390/ijms13032618
dc.identifier.issn14220067
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/110211
dc.description.abstractDomain III of the dengue virus envelope protein (EDIII, aa295-395) has an immunoglobulin fold and is the proposed receptor-binding domain of the virus. Previous studies have shown that monoclonal antibodies against EDIII can be neutralizing and have therapeutic potential. Here, cloned Fab-phage libraries of human and mouse origin were screened for DENV specific antibodies. Firstly, bacterially expressed EDIII or whole virus particles were used as bait in biopanning against a large naïve human Fab-phage library (>10 billion independent clones). Multiple panning strategies were employed, and in excess of 1000 clones were screened, but all of the antibodies identified bound the envelope in regions outside EDIII suggesting EDIII antibodies are virtually absent from the naïve human repertoire. Next, a chimeric Fab-phage library was constructed from a panel of EDIII specific mouse hybridomas by pooling the VH and VL chain sequences from the hybridomas and cloning these into the pComb3X phagemid vector with human CH and CL encoding sequences. Biopanning against EDIII identified a unique antibody (C9) that cross-reacts with EDIII from DENV1-3 and, in the IgG format, binds and neutralizes DENV2 in cell-based assays. Sequence analysis and saturation mutagenesis of complementary determining regions (CDR) in the C9 light chain suggest an antigen recognition model in which the LCDR3 is a key determinant of EDIII specificity, while modifications in LCDR1 and LCDR2 affect DENV serotype cross-reactivity. Overall, this study supports the current prevailing opinion that neutralizing anti-EDIII monoclonal antibodies can be readily generated in murine systems, but in humans the anti-DENV immune response is directed away from domain III. © 2012 by the authors.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.3390/ijms13032618
dc.sourceScopus
dc.subjectDengue
dc.subjectDomain III
dc.subjectE protein
dc.subjectFab
dc.subjectHybridoma
dc.subjectPhage display
dc.typeArticle
dc.contributor.departmentDUKE-NUS GRADUATE MEDICAL SCHOOL S'PORE
dc.description.doi10.3390/ijms13032618
dc.description.sourcetitleInternational Journal of Molecular Sciences
dc.description.volume13
dc.description.issue3
dc.description.page2618-2635
dc.identifier.isiut000302174500006
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