- EMDB-27352: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclo... -
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データベース: EMDB / ID: EMD-27352
タイトル
nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
マップデータ
nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
試料
複合体: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
AI136621
米国
Bill & Melinda Gates Foundation
INV-004923
米国
Bill & Melinda Gates Foundation
INV-002916
米国
引用
ジャーナル: Cell Rep Methods / 年: 2023 タイトル: Increasing sensitivity of antibody-antigen interactions using photo-cross-linking. 著者: Alba Torrents de la Peña / Leigh M Sewall / Rebeca de Paiva Froes Rocha / Abigail M Jackson / Payal P Pratap / Sandhya Bangaru / Christopher A Cottrell / Subhasis Mohanty / Albert C Shaw / Andrew B Ward / 要旨: Understanding antibody-antigen interactions in a polyclonal immune response in humans and animal models is critical for rational vaccine design. Current approaches typically characterize antibodies ...Understanding antibody-antigen interactions in a polyclonal immune response in humans and animal models is critical for rational vaccine design. Current approaches typically characterize antibodies that are functionally relevant or highly abundant. Here, we use photo-cross-linking and single-particle electron microscopy to increase antibody detection and unveil epitopes of low-affinity and low-abundance antibodies, leading to a broader structural characterization of polyclonal immune responses. We employed this approach across three different viral glycoproteins and showed increased sensitivity of detection relative to currently used methods. Results were most noticeable in early and late time points of a polyclonal immune response. Additionally, the use of photo-cross-linking revealed intermediate antibody binding states and demonstrated a distinctive way to study antibody binding mechanisms. This technique can be used to structurally characterize the landscape of a polyclonal immune response of patients in vaccination or post-infection studies at early time points, allowing for rapid iterative design of vaccine immunogens.
nsEM half map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
nsEM half map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
全体 : nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclo...
全体
名称: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
要素
複合体: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
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超分子 #1: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclo...
超分子
名称: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker タイプ: complex / ID: 1 / 親要素: 0 詳細: HA recombinantly expressed in HEK 293F cells Fab samples isolated from several individuals at different timepoints
15 ug of H1 A/Mich15/045/15 was complexed overnight with 500 ug of polyclonal Fab samples and photo-cross-linked. Sample was SEC purified and the complex peak was concentrated to ~0.02 mg/ml and imaged