Immunological Data Discovery Index
Immunological Data Discovery Index
| authorizations: |
registration not required
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| accessURL: |
http://www.iedb.org/reference/1027139 |
| landingPage: |
http://www.iedb.org/assay/2021443 |
| type: |
Submission
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| dates: |
2014
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| study type: | b cell assays |
| subject species: | Dengue virus 3 |
| fullName: |
Benjamin J. Doranz Ph.D.
Kristen M. Kahle Ph.D.
Jennifer M. Pfaff
Sherri L. Swayne
Xiaoxiao Xiang Ph.D.
Scott A. Smith MD, Ph.D
James E. Crowe, Jr. MD
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| method: |
microarray
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| name: |
Mapping Antibody Epitopes and Functional Regions of Dengue Envelope Proteins
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| description: |
1E23 binds the DENV prM region.
To obtain epitope maps at a resolution of individual amino acids for monoclonal antibodies (MAbs) directed against the immunodominant envelope protein prM/E of Dengue virus (DENV), we used a novel Shotgun Mutagenesis approach (www.integralmolecular.com; Paes et al., 2009 JACS 131: 6952-6954). Shotgun Mutagenesis provided a residue-by-residue evaluation of the interaction between the prM/E target protein and anti-DENV MAbs. A comprehensive library of mutated DENV-3 (CH53489 strain) prM/E expression plasmids (1440 individual mutants) was expressed in human cells and subjected to high-throughput analyses of antibody reactivity using microplate-based immunofluorescence assays. We have screened human anti-DENV MAbs, derived from naturally infected patients, and murine MAbs, derived from DENV reporter virus immunizations (Mattia et al., 2011 PLoS ONE 6 (11): e27252). Mutations were identified as critical to a specific MAb epitope if they did not support reactivity of the MAb, but did support reactivity of other conformation-dependent MAbs. This counter-screen strategy facilitated the exclusion of prM/E mutants that were globally or locally misfolded, or that had expression defects.
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| name: |
iedb
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| homePage: |
http://www.iedb.org |
