Construction of single-chain Fv with two possible CDR3H conformations but similar inter-molecular forces that neutralize bovine herpesvirus 1
dc.contributor.author | Koti, M. | |
dc.contributor.author | Farrugia, W. | |
dc.contributor.author | Nagy, E. | |
dc.contributor.author | Ramsland, Paul | |
dc.contributor.author | Kaushik, A. | |
dc.date.accessioned | 2017-01-30T11:10:16Z | |
dc.date.available | 2017-01-30T11:10:16Z | |
dc.date.created | 2016-09-12T08:36:44Z | |
dc.date.issued | 2010 | |
dc.identifier.citation | Koti, M. and Farrugia, W. and Nagy, E. and Ramsland, P. and Kaushik, A. 2010. Construction of single-chain Fv with two possible CDR3H conformations but similar inter-molecular forces that neutralize bovine herpesvirus 1. Molecular Immunology. 47 (5): pp. 953-960. | |
dc.identifier.uri | http://hdl.handle.net/20.500.11937/9046 | |
dc.identifier.doi | 10.1016/j.molimm.2009.11.011 | |
dc.description.abstract |
Bovine herpesvirus 1 (BoHV-1) causes respiratory and genital diseases in cattle for which available vaccines do not confer adequate protection. Since passive immunization with antibodies permits disease prevention, single-chain fragment variable (scFv), originating from a monoclonal bovine IgG1 antibody against BoHV-1, were constructed and expressed in Pichia pastoris in V?-VH orientation via a flexible seven-amino acid linker. Similar to the intact IgG, the purified recombinant scFv neutralized BoHV-1 in vitro and recognized viral antigens in BoHV-1 infected MDBK cells by immunofluorescence. Homology modeling of the Fv predicts two distinct conformations for CDR3H. Firstly, a long protruding CDR3H conformation where no disulfide linkage occurred between two "non-canonical" Cys residues resulted in a large binding cavity between V? and VH. Secondly, a smaller potential antigen-binding cavity is predicted with a disulfide linkage between the two Cys residues of CDR3H creating a six-membered loop in the ascending polypeptide, which fitted into the space between V? and VH. Despite such potential configurational diversity of the antigen-binding site, the electrostatic surface potentials that would interact with the BoHV-1 epitope are largely similar for both the topographies where salt-bridge type electrostatic interactions likely occur at the edges of the binding site. Given that IgG1 antibody against BoHV-1 is clonally selected, it is likely that disulfide-stabilized broader and flatter surface topography is specifically generated to accommodate the predicted carbohydrate neutralizing B-epitope on BoHV-1. The specificity and neutralizing capacity for BoHV-1 of the scFv should make this bovine antibody fragment a useful diagnostic and potential therapeutic candidate for an important viral pathogen in cattle. © 2009 Elsevier Ltd. All rights reserved. | |
dc.title | Construction of single-chain Fv with two possible CDR3H conformations but similar inter-molecular forces that neutralize bovine herpesvirus 1 | |
dc.type | Journal Article | |
dcterms.source.volume | 47 | |
dcterms.source.number | 5 | |
dcterms.source.startPage | 953 | |
dcterms.source.endPage | 960 | |
dcterms.source.issn | 0161-5890 | |
dcterms.source.title | Molecular Immunology | |
curtin.department | School of Biomedical Sciences | |
curtin.accessStatus | Fulltext not available |
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