ジャーナル: Proc Natl Acad Sci U S A / 年: 2014 タイトル: Structures of protective antibodies reveal sites of vulnerability on Ebola virus. 著者: Charles D Murin / Marnie L Fusco / Zachary A Bornholdt / Xiangguo Qiu / Gene G Olinger / Larry Zeitlin / Gary P Kobinger / Andrew B Ward / Erica Ollmann Saphire / 要旨: Ebola virus (EBOV) and related filoviruses cause severe hemorrhagic fever, with up to 90% lethality, and no treatments are approved for human use. Multiple recent outbreaks of EBOV and the likelihood ...Ebola virus (EBOV) and related filoviruses cause severe hemorrhagic fever, with up to 90% lethality, and no treatments are approved for human use. Multiple recent outbreaks of EBOV and the likelihood of future human exposure highlight the need for pre- and postexposure treatments. Monoclonal antibody (mAb) cocktails are particularly attractive candidates due to their proven postexposure efficacy in nonhuman primate models of EBOV infection. Two candidate cocktails, MB-003 and ZMAb, have been extensively evaluated in both in vitro and in vivo studies. Recently, these two therapeutics have been combined into a new cocktail named ZMapp, which showed increased efficacy and has been given compassionately to some human patients. Epitope information and mechanism of action are currently unknown for most of the component mAbs. Here we provide single-particle EM reconstructions of every mAb in the ZMapp cocktail, as well as additional antibodies from MB-003 and ZMAb. Our results illuminate key and recurring sites of vulnerability on the EBOV glycoprotein and provide a structural rationale for the efficacy of ZMapp. Interestingly, two of its components recognize overlapping epitopes and compete with each other for binding. Going forward, this work now provides a basis for strategic selection of next-generation antibody cocktails against Ebola and related viruses and a model for predicting the impact of ZMapp on potential escape mutations in ongoing or future Ebola outbreaks.
全体 : Fab fragment of c2G4 chimerized human monoclonal IgG1 antibody bo...
全体
名称: Fab fragment of c2G4 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein
要素
試料: Fab fragment of c2G4 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein
タンパク質・ペプチド: Ebola virus glycoprotein
タンパク質・ペプチド: chimerized human IgG1 antigen binding fragment c2G4
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超分子 #1000: Fab fragment of c2G4 chimerized human monoclonal IgG1 antibody bo...
超分子
名称: Fab fragment of c2G4 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein タイプ: sample / ID: 1000 / 集合状態: Three Fab fragments bound to a single GP / Number unique components: 2
分子 #2: chimerized human IgG1 antigen binding fragment c2G4
分子
名称: chimerized human IgG1 antigen binding fragment c2G4 / タイプ: protein_or_peptide / ID: 2 / Name.synonym: c2G4 Fab / コピー数: 3 / 集合状態: monomer / 組換発現: Yes
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: human
組換発現
生物種: Nicotiana benthamiana (ナス科)
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実験情報
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構造解析
手法
ネガティブ染色法
解析
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
0.03 mg/mL
緩衝液
pH: 7.4 / 詳細: 20 mM Tris, 150 mM NaCl
染色
タイプ: NEGATIVE 詳細: To prepare negative stain grids, a 4 uL aliquot of each complex, which had been diluted to a concentration of ~0.03 ug/mL with TBS buffer, was placed for 15 seconds onto carbon-coated 400 Cu ...詳細: To prepare negative stain grids, a 4 uL aliquot of each complex, which had been diluted to a concentration of ~0.03 ug/mL with TBS buffer, was placed for 15 seconds onto carbon-coated 400 Cu mesh grids that had been plasma cleaned for 20 s (Gatan), blotted off on the edge of the grid, then immediately stained for 30 s with 4 uL of 2% uranyl formate. The stain was blotted off on the edge of the grid and the grid was allowed to dry.