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基本情報
登録情報 | データベース: EMDB / ID: EMD-7555 | |||||||||
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タイトル | CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 | |||||||||
![]() | BG505 SOSIP.664 in complex with serum from rabbit 3417 post boost 1. Map of antibodies bound to trimer after several rounds of 3D classification in CryoSparc. | |||||||||
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生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.7 Å | |||||||||
![]() | Turner HL / Ward AB / Nogal B | |||||||||
![]() | ![]() タイトル: Electron-Microscopy-Based Epitope Mapping Defines Specificities of Polyclonal Antibodies Elicited during HIV-1 BG505 Envelope Trimer Immunization. 著者: 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 ...著者: 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 / ![]() ![]() 要旨: 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. | |||||||||
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 96.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 11.3 KB 11.3 KB | 表示 表示 | ![]() |
画像 | ![]() | 65.8 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 78 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 77.1 KB | 表示 | |
XML形式データ | ![]() | 493 B | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 7552C ![]() 7553C ![]() 7554C ![]() 7556C ![]() 7557C ![]() 7570C ![]() 7887C ![]() 7888C ![]() 7889C ![]() 7890C ![]() 7891C ![]() 7892C ![]() 7893C ![]() 7894C ![]() 7895C ![]() 7896C ![]() 7903C ![]() 7904C ![]() 7906C ![]() 6cjkC ![]() 6didC C: 同じ文献を引用 ( |
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ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | BG505 SOSIP.664 in complex with serum from rabbit 3417 post boost 1. Map of antibodies bound to trimer after several rounds of 3D classification in CryoSparc. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.15 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
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試料の構成要素
-全体 : Complex of BG505 SOSIP.664 and serum antibodies from post boost 1...
全体 | 名称: Complex of BG505 SOSIP.664 and serum antibodies from post boost 1 from rabbit 3417. Subset of particles from dataset. |
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要素 |
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-超分子 #1: Complex of BG505 SOSIP.664 and serum antibodies from post boost 1...
超分子 | 名称: Complex of BG505 SOSIP.664 and serum antibodies from post boost 1 from rabbit 3417. Subset of particles from dataset. タイプ: complex / ID: 1 / 親要素: 0 詳細: Serum was purified through SEC after using a Protein A or G column to capture complex then cleave IgG releasing Fab and trimer. Substoichiometric occupancy and flexibility show lack of ...詳細: Serum was purified through SEC after using a Protein A or G column to capture complex then cleave IgG releasing Fab and trimer. Substoichiometric occupancy and flexibility show lack of density in Fab region while trimer core is refined to higher resolution. Map shows strong density of single Fab bound to glycan hole and low densities of Fabs bound to the trimer base. |
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由来(天然) | 生物種: ![]() ![]() |
組換発現 | 生物種: ![]() |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 1.44 mg/mL |
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緩衝液 | pH: 7.4 / 詳細: TBS |
グリッド | モデル: Quantifoil R1.2/1.3 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK IV |
詳細 | Sample in TBS, added 0.5uL of DDM at 0.42mM. Sample was monodisperse |
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電子顕微鏡法
顕微鏡 | FEI TECNAI ARCTICA |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - 画像ごとのフレーム数: 1-46 / 平均露光時間: 11.5 sec. / 平均電子線量: 60.0 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Talos Arctica / 画像提供: FEI Company |