{"id":2597,"date":"2020-11-02T16:37:48","date_gmt":"2020-11-02T16:37:48","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/?p=2597"},"modified":"2024-02-14T23:58:02","modified_gmt":"2024-02-14T23:58:02","slug":"developing-analytics-to-move-gene-therapy-forward","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/developing-analytics-to-move-gene-therapy-forward\/","title":{"rendered":"Developing Analytics to Move Gene Therapy Forward"},"content":{"rendered":"<p class=\"lia-message-body-content\">With hundreds of gene therapy clinical trials ongoing and about a dozen gene therapy products already on the market, we must provide scientists with the tools needed to characterize this new generation of biotherapeutics. Traditional biologics, such as monoclonal antibodies, are manufactured by expressing one protein in a cell line and purifying it.<\/p>\n<p class=\"lia-message-body-content\"><span class=\"lia-inline-image-display-wrapper\"><img decoding=\"async\" class=\"lia-media-image alignright\" title=\"virus_110220\" role=\"button\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/Developing-Analytics-to-Move-Gene-Therapy-Forward-0.jpg\" alt=\"virus_110220\" \/><\/span>For gene therapy products, viruses, like the adeno-associated virus (AAV), are typically incorporated into the final product as the gene delivery vehicle. With this addition, AAV attributes need to be added to the product and process monitoring toolbox, including AAV purity, identity, and empty and full capsid analysis. With the right analytical tools for complex gene therapy characterization and monitoring through the development process, scientists can more quickly develop safe, high-quality gene therapy products while reducing time-to-market and reducing costs of manufacturing.<\/p>\n<h3 id=\"toc-hId--1902364986\"><strong>Empty\/ Partial\/ Full Capsid Analysis<\/strong><\/h3>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"size-full wp-image-12342 alignright\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage_110220-1.jpg\" alt=\"\" width=\"300\" height=\"166\" \/><\/p>\n<p>AAV vectors are common gene delivery vehicles. Empty capsids, which do not contain the gene therapy of interest, and partial capsids, those containing only a fragment of the gene of interest, are by-products of the AAV production and can impact product safety and efficacy. The amount of full, partial, and empty capsids, therefore, needs to be characterized and monitored through process development. There are various methods used to characterize full, partial, and empty capsid levels including analytical ultracentrifugation, transmission electron microscopy, charge detection mass spectrometry, and anion exchange chromatography.<\/p>\n<p><strong>Methods in Action:<\/strong> In the upcoming webinar (November 11, 2020) by Julia Baek titled, \u201c<a href=\"https:\/\/event.on24.com\/eventRegistration\/EventLobbyServlet?target=reg20.jsp&amp;referrer=&amp;eventid=2222321&amp;sessionid=1&amp;key=320A029BB82F55EF2B8D2D8750F1D61C&amp;regTag=841363&amp;sourcepage=register\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Developing analytics to support the next generation of gene therapies<\/a>\u201d, Julia will discuss the development of fast <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/054997#\/054997\" target=\"_blank\" rel=\"nofollow noopener\">anion exchange chromatography<\/a> methods for the analysis of full and empty AAV capsids. Baseline separation of full and empty capsids of several different serotypes using either salt gradient or pH gradient methods will be presented.<\/p>\n<p><a href=\"https:\/\/event.on24.com\/wcc\/r\/2222321\/320A029BB82F55EF2B8D2D8750F1D61C\/841363?partnerref=E.20CMD.PB103.31533.01\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Register for this or any of our Leaders in BioPharma Characterization and Control webinars here<\/a>.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter size-full wp-image-12344\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage2_110220-1.jpg\" alt=\"\" width=\"853\" height=\"427\" srcset=\"https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage2_110220-1.jpg 853w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage2_110220-1-300x150.jpg 300w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage2_110220-1-768x384.jpg 768w\" sizes=\"auto, (max-width: 853px) 100vw, 853px\" \/><\/p>\n<h3 id=\"toc-hId--1873735835\"><strong>Capsid Protein Characterization<\/strong><\/h3>\n<p>It is important to understand the capsid identity because each AAV serotype has unique capsid proteins that transfer the genetic material to specific cells or organs. Depending on the therapeutic target area, the correct AAV serotype is chosen and identity and purity are monitored. All AAV capsids consist of three proteins (VP1, VP2, and VP3) that share high-sequence homology, and ensuring the identity and purity of these proteins are critical to viral infectivity and viral transfer. Given the importance of the serotype and the capsid protein composition, there exists a need for methods to identify and monitor the capsid and the capsid proteins through gene therapy development and manufacturing.<\/p>\n<p><strong>Methods in Action:<\/strong> In a paper by <a href=\"https:\/\/www.sciencedirect.com\/science\/article\/pii\/S0731708520313674\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Liu et al<\/a>, capsid proteins from eight AAV serotypes were characterized using HILIC chromatography coupled to a <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/IQLAAEGAAPFALGMBDK?gclid=Cj0KCQjw5eX7BRDQARIsAMhYLP8eWmGcsVzmNyScNecvflL66eyBehTSMwbH7_6zOHdAmrGFQNs_YF4aAmhlEALw_wcB&amp;ce=E.20CMD.DL102.29343.01&amp;cid=E.20CMD.DL102.29343.01&amp;ef_id=Cj0KCQjw5eX7BRDQARIsAMhYLP8eWmGcsVzmNyScNecvflL66eyBehTSMwbH7_6zOHdAmrGFQNs_YF4aAmhlEALw_wcB:G:s&amp;s_kwcid=AL!3652!3!249703668935!e!!g!!q%20exactive%20plus#\/IQLAAEGAAPFALGMBDK?gclid=Cj0KCQjw5eX7BRDQARIsAMhYLP8eWmGcsVzmNyScNecvflL66eyBehTSMwbH7_6zOHdAmrGFQNs_YF4aAmhlEALw_wcB&amp;ce=E.20CMD.DL102.29343.01&amp;cid=E.20CMD.DL102.29343.01&amp;ef_id=Cj0KCQjw5eX7BRDQARIsAMhYLP8eWmGcsVzmNyScNecvflL66eyBehTSMwbH7_6zOHdAmrGFQNs_Y\" target=\"_blank\" rel=\"nofollow noopener\">mass spectrometer<\/a>.<\/p>\n<p>In the upcoming webinar (November 11, 2020) by Julia Baek and Estee Toole titled, \u201c<a href=\"https:\/\/event.on24.com\/eventRegistration\/EventLobbyServlet?target=reg20.jsp&amp;referrer=&amp;eventid=2222321&amp;sessionid=1&amp;key=320A029BB82F55EF2B8D2D8750F1D61C&amp;regTag=841363&amp;sourcepage=register\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Developing analytics to support the next generation of gene therapies<\/a>\u201d, Estee will discuss her work on capsid protein characterization by peptide mapping.<\/p>\n<h3 id=\"toc-hId--1845106684\"><strong>Intact Capsid Mass Analysis<\/strong><\/h3>\n<p>Monitoring intact capsids allows for mass confirmation of empty versus genome packed capsids. To add to the complexity, genome loaded viral vector particles are even more heterogeneous because the package can be a mix of genomic material. Using charge-detection mass spectrometry (CD-MS) opens the door to analyzing these macromolecules with high mass heterogeneity, providing a means to quality control genome packaging in production of gene therapy products.<\/p>\n<p><strong>Methods in Action:<\/strong> In a paper by <a href=\"https:\/\/www.biorxiv.org\/content\/10.1101\/717413v1\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Worner et al<\/a>, Orbitrap-based CD-MS was used to successfully resolve distinct distributions of empty and full AAV8 particles.<\/p>\n<h3 id=\"toc-hId--1816477533\"><strong>More Resources<\/strong><\/h3>\n<p><a href=\"https:\/\/event.on24.com\/wcc\/r\/2222321\/320A029BB82F55EF2B8D2D8750F1D61C\/841363?partnerref=E.20CMD.PB103.31533.01\" target=\"_blank\" rel=\"nofollow noopener noreferrer\">Register for any of our Leaders in BioPharma Characterization and Control webinars here<\/a><\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter size-full wp-image-12345\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage3_110220-1.jpg\" alt=\"\" width=\"990\" height=\"212\" srcset=\"https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage3_110220-1.jpg 990w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage3_110220-1-300x64.jpg 300w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/blogimage3_110220-1-768x164.jpg 768w\" sizes=\"auto, (max-width: 990px) 100vw, 990px\" \/><\/p>\n<p>Visit site for information on <a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/industrial\/mass-spectrometry\/liquid-chromatography-mass-spectrometry-lc-ms\/lc-ms-systems\/orbitrap-lc-ms\/q-exactive-orbitrap-mass-spectrometers\/q-exactive-uhmr.html\" target=\"_blank\" rel=\"nofollow noopener\">Thermo Scientific<sup>TM<\/sup> Q Exactive<sup>TM<\/sup> UHMR Hybrid Quadrupole-Orbitrap<sup>TM<\/sup> mass spectrometer<\/a>.<\/p>\n<p><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/clinical\/cell-gene-therapy.htm\" target=\"_blank\" rel=\"nofollow noopener\">Visit for Cell and Gene Therapy production solutions<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>With hundreds of gene therapy clinical trials ongoing and about a dozen gene therapy products already on the market, we must provide scientists with<\/p>\n","protected":false},"author":1327,"featured_media":2599,"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_memberships_contains_paid_content":false,"footnotes":""},"categories":[38],"tags":[41],"division":[],"class_list":{"0":"post-2597","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-biopharmaceutical","8":"tag-biopharmaceutical","9":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- 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