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Yorodumi- EMDB-7906: Negative stain EM map of BG505 SOSIP.644 in complex with PG9 Fab ... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-7906 | |||||||||
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Title | Negative stain EM map of BG505 SOSIP.644 in complex with PG9 Fab and serum from rabbit 3417 PB1 | |||||||||
Map data | Negative stain EM map of BG505 SOSIP.664 in complex with PG9 Fab and serum Fab from rabbit 3417 PB1 | |||||||||
Sample |
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Biological species | Human immunodeficiency virus | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 18.45 Å | |||||||||
Authors | Turner HL / Ward AB / Nogal B | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Immunity / Year: 2018 Title: Electron-Microscopy-Based Epitope Mapping Defines Specificities of Polyclonal Antibodies Elicited during HIV-1 BG505 Envelope Trimer Immunization. Authors: Matteo Bianchi / Hannah L Turner / Bartek Nogal / Christopher A Cottrell / David Oyen / Matthias Pauthner / Raiza Bastidas / Rebecca Nedellec / Laura E McCoy / Ian A Wilson / Dennis R Burton ...Authors: Matteo Bianchi / Hannah L Turner / Bartek Nogal / Christopher A Cottrell / David Oyen / Matthias Pauthner / Raiza Bastidas / Rebecca Nedellec / Laura E McCoy / Ian A Wilson / Dennis R Burton / Andrew B Ward / Lars Hangartner / Abstract: Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a ...Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a barrier to fully understanding the humoral response to antigens and hinders rational vaccine design efforts. Here, we describe a method of characterizing polyclonal responses by using electron microscopy, and we applied this method to the immunization of rabbits with an HIV-1 envelope glycoprotein vaccine candidate, BG505 SOSIP.664. We detected known epitopes within the polyclonal sera and revealed how antibody responses evolved during the prime-boosting strategy to ultimately result in a neutralizing antibody response. We uncovered previously unidentified epitopes, including an epitope proximal to one recognized by human broadly neutralizing antibodies as well as potentially distracting non-neutralizing epitopes. Our method provides an efficient and semiquantitative map of epitopes that are targeted in a polyclonal antibody response and should be of widespread utility in vaccine and infection studies. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_7906.map.gz | 6 MB | EMDB map data format | |
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Header (meta data) | emd-7906-v30.xml emd-7906.xml | 11.7 KB 11.7 KB | Display Display | EMDB header |
Images | emd_7906.png | 30.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-7906 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-7906 | HTTPS FTP |
-Validation report
Summary document | emd_7906_validation.pdf.gz | 298 KB | Display | EMDB validaton report |
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Full document | emd_7906_full_validation.pdf.gz | 297.6 KB | Display | |
Data in XML | emd_7906_validation.xml.gz | 5.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7906 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7906 | HTTPS FTP |
-Related structure data
Related structure data | 7552C 7553C 7554C 7555C 7556C 7557C 7570C 7887C 7888C 7889C 7890C 7891C 7892C 7893C 7894C 7895C 7896C 7903C 7904C 6cjkC 6didC C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_7906.map.gz / Format: CCP4 / Size: 11.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Negative stain EM map of BG505 SOSIP.664 in complex with PG9 Fab and serum Fab from rabbit 3417 PB1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.05 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Complex of BG505 SOSIP.664 with PG9 Fab and serum from rabbit 3417 PB1
Entire | Name: Complex of BG505 SOSIP.664 with PG9 Fab and serum from rabbit 3417 PB1 |
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Components |
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-Supramolecule #1: Complex of BG505 SOSIP.664 with PG9 Fab and serum from rabbit 3417 PB1
Supramolecule | Name: Complex of BG505 SOSIP.664 with PG9 Fab and serum from rabbit 3417 PB1 type: complex / ID: 1 / Parent: 0 Details: Complex was purified through SEC after adding excess PG9 and serum |
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Source (natural) | Organism: Human immunodeficiency virus |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant strain: HEK293F |
-Experimental details
-Structure determination
Method | negative staining |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.114 mg/mL |
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Buffer | pH: 7.4 / Details: TBS |
Staining | Type: NEGATIVE / Material: Uranyl Formate / Details: at 2% |
Grid | Model: Homemade / Material: COPPER / Mesh: 400 / Support film - Material: CELLULOSE ACETATE / Pretreatment - Type: GLOW DISCHARGE |
Details | Sample in TBS, stained with UF |
-Electron microscopy
Microscope | FEI TECNAI SPIRIT |
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Image recording | Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-46 / Number grids imaged: 1 / Number real images: 438 / Average exposure time: 0.5 sec. / Average electron dose: 25.0 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: LAB6 |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: SIDE ENTRY, EUCENTRIC |
Experimental equipment | Model: Tecnai Spirit / Image courtesy: FEI Company |