{"id":652,"date":"2022-02-24T05:10:05","date_gmt":"2022-02-24T05:10:05","guid":{"rendered":"https:\/\/admin.acceleratingscience.com\/atomic-resolution\/2022\/02\/24\/cryo-em-epitope-mapping-drug-discovery\/"},"modified":"2022-06-07T17:20:50","modified_gmt":"2022-06-07T17:20:50","slug":"cryo-em-epitope-mapping-drug-discovery","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/atomic-resolution\/cryo-em-epitope-mapping-drug-discovery\/","title":{"rendered":"Rapid Cryo-EM Epitope Mapping Could Enhance Antibody Drug Discovery"},"content":{"rendered":"<p>Historically, cryo-electron microscopy (cryo-EM) structure determination required operators to collect one dataset after another. But with the onset of the SARS-CoV-2 pandemic and a subsequent queue of variants continuously impacting our ability to prevent infection with existing solutions, there\u2019s a clear need for faster, more efficient, data collection for the purpose of drug discovery. In collaboration with Takeda Pharmaceuticals and Utrecht University, we demonstrate the power of high-throughput cryo-EM for epitope mapping, including the potential for improved antibody therapeutics.<\/p>\n<hr \/>\n<p><a href=\"https:\/\/ter.li\/ftyhky\"><em>Read our whitepaper: High-throughput cryo-EM epitope mapping of SARS-CoV-2 spike protein antibodies using EPU Multigrid<\/em><\/a><em> &gt;&gt;<\/em><\/p>\n<hr \/>\n<h2 style=\"font-size: 100%\"><strong>Cryo-EM for rapid epitope mapping<\/strong><\/h2>\n<p>An epitope is a part of a molecule against which an antibody attaches itself.<\/p>\n<p>Therapeutic antibodies targeting the SARS-CoV-2 spike protein can play an important role in reducing the burden of the pandemic by protecting vulnerable populations against severe and potentially life-threatening disease.<\/p>\n<p>Epitope mapping of large antibody panels allows researchers to identify vulnerable sites on the spike protein, predict and interpret the effect of new mutations, and speed decision-making in selecting antibody combinations that target non-overlapping epitopes. Moreover, structural information about epitopes can help identify liabilities in drug discovery pipelines in response to new emerging variants.<\/p>\n<p>Therefore, in <a href=\"https:\/\/ter.li\/ftyhky\">collaboration with\u00a0Takeda Pharmaceuticals and\u00a0Utrecht University<\/a>, we set out to determine whether cryo-EM permits rapid epitope mapping for a large antibody panel.<\/p>\n<p>Our study launched in the wake of the Omicron variant and subsequent strains, which escape most neutralizing antibodies, as well as the realization that we need additional tools for long-lasting prophylaxis against\u00a0SARS-COV-2\u00a0infections.<\/p>\n<h2 style=\"font-size: 100%\"><strong>Newfound speed, quality, and automation of cryo-EM data collection<\/strong><\/h2>\n<p>Robust grid preparation and screening are critical for determining structures from every grid, and successful grid preparation is intimately linked to protein quality and concentration.<\/p>\n<p>To evaluate the robustness of our spike protein sample, and to ultimately gauge the suitability of our 12 complexes for structural determination, we used the new automation capabilities of Thermo Scientific <a href=\"https:\/\/ter.li\/a3xk9r\">EPU Multigrid software<\/a> on our <a href=\"https:\/\/ter.li\/8xcuww\">Glacios Cryo-TEM<\/a>.<\/p>\n<p>The twelve screened grids were then subjected to unattended EPU Multigrid data collection using our <a href=\"https:\/\/ter.li\/hlgibq\">Krios G4 Cryo-TEM<\/a> equipped with an E-CFEG, <a href=\"https:\/\/ter.li\/xlgzmw\">Selectris X Energy Filter<\/a>, and <a href=\"https:\/\/ter.li\/1r7ft0\">Falcon 4 Direct Electron Detector<\/a>.<\/p>\n<div id=\"attachment_2165\" style=\"width: 770px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-2165\" class=\"wp-image-2165 size-large\" title=\"High-throughput cryo-EM epitope mapping of SARS-CoV-2 spike protein antibodies\" src=\"http:\/\/admin.acceleratingscience.com\/atomic-resolution\/wp-content\/uploads\/sites\/22\/2022\/04\/Takeda-UU-WP_blog1-003-1-1024x420.png\" alt=\"High-throughput cryo-EM epitope mapping of SARS-CoV-2 spike protein antibodies\" width=\"760\" height=\"312\" \/><p id=\"caption-attachment-2165\" class=\"wp-caption-text\">High-throughput cryo-EM epitope mapping of SARS-CoV-2 spike protein antibodies. Spike-Fab complex grids were imaged using Krios G4 cryo-TEM and EPU Multigrid, which resulted in twelve sub 3 \u00c5 cryo-EM reconstructions allowing epitope mapping and classification.<\/p><\/div>\n<p>In a single 48 hour-long session, we set EPU Multigrid to collect 1,000 movies per grid, achieving a speed of 250 movies per hour.<\/p>\n<p>This workflow enabled us to obtain 12 sub-3-angstrom structures of SARS-CoV-2 spike protein with different complexes and to determine their epitopes. Our results show the significant advances made in speed, quality, and automation of cryo-EM data collection.<\/p>\n<p><em>&#8220;These exciting results show how state-of-the-art technology, and an optimized sample preparation workflow, can be used to accelerate antibody drug discovery, which is of great importance for COVID-19 and future pandemic threats,\u201d <\/em>said collaborator Daniel Hurdiss, Assistant Professor at Utrecht University.<\/p>\n<p><strong><a href=\"https:\/\/ter.li\/ftyhky\">Get an in-depth look at this workflow &gt;&gt;<\/a><\/strong><\/p>\n<p>We believe cryo-EM\u2019s high-resolution revolution will soon be followed by a high-throughput revolution, which will improve existing applications through increased efficiency and reduced cost per dataset. It will also extend the range of cryo-EM applications to include fragment-based drug discovery and epitope mapping of large antibody panels.<\/p>\n<p>\/\/<\/p>\n<p><em>Mazdak <\/em><em>Radjainia is a Senior Staff Scientist at Thermo Fisher Scientific<\/em><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Historically, cryo-electron microscopy (cryo-EM) structure determination required operators to collect one dataset after another. But with the onset of the SARS-CoV-2 pandemic and a subsequent queue of variants continuously impacting our ability to prevent infection with existing solutions, there\u2019s a clear need for faster, more efficient, data collection for the purpose of drug discovery. In<\/p>\n","protected":false},"author":1115,"featured_media":654,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_feature_clip_id":0,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_post_was_ever_published":false},"categories":[24,51,33],"tags":[15,16,117],"division":[],"class_list":{"0":"post-652","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-cryo-electron-microscopy-cryo-em","8":"category-customer-spotlights","9":"category-general","10":"tag-cryo-electron-microscopy","11":"tag-cryo-em","12":"tag-drug-discovery","13":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- 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