{"id":661,"date":"2019-07-08T09:51:46","date_gmt":"2019-07-08T09:51:46","guid":{"rendered":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/?p=661"},"modified":"2019-07-08T09:51:46","modified_gmt":"2019-07-08T09:51:46","slug":"hybrid-mass-spectrometry-methods-reveal-lot-tolot-differences-and-delineate-the-effects-of-glycosylation-on-the-tertiary-structure-of-herceptin","status":"publish","type":"post","link":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/2019\/07\/08\/hybrid-mass-spectrometry-methods-reveal-lot-tolot-differences-and-delineate-the-effects-of-glycosylation-on-the-tertiary-structure-of-herceptin\/","title":{"rendered":"Hybrid mass spectrometry methods reveal lot-tolot differences and delineate the effects of glycosylation on the tertiary structure of Herceptin"},"content":{"rendered":"

To quantify the measurable variations in the structure of a biopharmaceutical product we systematically
\nevaluate three lots of Herceptin\u00ae, two mAb standards and an intact Fc-hinge fragment. Each mAb is
\nexamined in three states; glycan intact, truncated (following endoS2 treatment) and fully deglycosylated.
\nDespite equivalence at the intact protein level, each lot of Herceptin\u00ae gives a distinctive signature in
\nthree different mass spectrometry approaches. Ion mobility mass spectrometry (IM-MS) shows that in
\nthe API, the attached N-glycans reduce the conformational spread of each mAb by 10.5\u201325%.
\nHydrogen\/deuterium exchange mass spectrometry (HDX-MS) data support this, with lower global
\ndeuterium uptake in solution when comparing intact to the fully deglycosylated protein. HDX-MS and
\nactivated IM-MS map the influence of glycans on the mAb and reveal allosteric effects which extend far
\nbeyond the Fc domains into the Fab region. Taken together, these findings and the supplied interactive
\ndata sets establish acceptance criteria with application for MS based characterisation of biosimilars and
\nnovel therapeutic mAbs.<\/p>\n

\"\"<\/p>\n

 <\/p>\n

Rosie Upton, a Lukasz G. Migas, a Kamila J. Pacholarz, a Richard G. Beniston,b<\/em>
\nSian Estdale, b David Firth b and Perdita E. Barran *a<\/em><\/p>\n

a Manchester Institute of Biotechnology, Michael Barber Centre for Collaborative Mass<\/em>
\nSpectrometry, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.<\/em>
\nE-mail: Perdita.barran@manchester.ac.uk<\/em>
\nb Covance Laboratories Ltd., Otley Road, Harrogate, HG3 1PY, UK<\/em><\/p>\n

https:\/\/pubs.rsc.org\/en\/content\/articlepdf\/2019\/sc\/c8sc05029e<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

To quantify the measurable variations in the structure of a biopharmaceutical product we systematically evaluate three lots of Herceptin\u00ae, two mAb standards and an intact Fc-hinge fragment. Each mAb is examined in three states; glycan intact, truncated (following endoS2 treatment) and fully deglycosylated. Despite equivalence at the intact protein level, each lot of Herceptin\u00ae gives … <\/p>\n

Continue reading<\/a><\/p>\n","protected":false},"author":2,"featured_media":662,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"jetpack_post_was_ever_published":false,"_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":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2}},"categories":[4,3],"tags":[],"class_list":["post-661","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-featured","category-publications","item-wrap"],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"https:\/\/i0.wp.com\/www.mbc.manchester.ac.uk\/barrangroup\/wp-content\/uploads\/sites\/2\/2019\/07\/c8sc05029e-f5_hi-res.gif?fit=1418%2C1218&ssl=1","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/p4Adwh-aF","_links":{"self":[{"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/posts\/661","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/comments?post=661"}],"version-history":[{"count":1,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/posts\/661\/revisions"}],"predecessor-version":[{"id":664,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/posts\/661\/revisions\/664"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/media\/662"}],"wp:attachment":[{"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/media?parent=661"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/categories?post=661"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.mbc.manchester.ac.uk\/barrangroup\/wp-json\/wp\/v2\/tags?post=661"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}