{"id":5590,"date":"2015-09-28T07:00:55","date_gmt":"2015-09-28T11:00:55","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/?p=5590"},"modified":"2016-04-29T16:52:56","modified_gmt":"2016-04-29T16:52:56","slug":"analytical-precision-for-developing-therapeutic-monoclonal-antibodies","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/proteomics\/analytical-precision-for-developing-therapeutic-monoclonal-antibodies\/","title":{"rendered":"Analytical Precision for Developing Therapeutic Monoclonal Antibodies"},"content":{"rendered":"<p><span class=\"thread\"><img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/admin.acceleratingscience.com\/wp-content\/uploads\/2015\/09\/cq5dam.thumbnail.250.250__1_1.png\" style=\"float: left;margin: 10px\" alt=\"Thermo Scientific Q Exactive\" width=\"250\" height=\"203\" \/>Monoclonal antibody (mAb) therapy<\/span>&nbsp;is used to to stimulate the immune system to defend itself against particular target cells. As researchers explore mAb treatments further, they are increasingly using mass spectrometry to selectively analyze&nbsp;human therapeutic mAbs for therapeutic development.<\/p>\n<p><span><span class=\"thread\"><span>According to Antwi et al., current mass spectrometry methods to analyze mAbs are limited and result in unreproducible data acquisition, a lack of analytical sensitivity and poor automation capabilities.<\/span><\/span><\/span>&nbsp;Therefore, the researchers describe a <a href=\"http:\/\/www.thermoscientific.com\/en\/products\/mass-spectrometric-immunoassay.html\" target=\"_blank\">MSIA (mass spectrometric immunoassay)<\/a> workflow (Thermo Scientific) that is better suited to&nbsp;therapeutic mAb pre-clinical discovery and development bioanalysis.&nbsp;MSIA technology combines the performance characteristics of traditional ligand binding with the benefits of mass spectrometry detection.<\/p>\n<p>Antwi et al. spiked mouse plasma with concentrations of&nbsp;adalimumab ranging from 20 to 8,000 g\/mL. They then loaded&nbsp;Streptavidin <a href=\"http:\/\/www.thermoscientific.com\/en\/product\/msia-pipette-tips.html\" target=\"_blank\">MSIA disposal automation research tips (<\/a><span class=\"thread\"><a href=\"http:\/\/www.thermoscientific.com\/en\/product\/msia-pipette-tips.html\" target=\"_blank\">D.A.R.T.s<\/a>)&nbsp;<\/span>with biotinylated <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/7103262100\" target=\"_blank\"><\/a><span class=\"thread\"><a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/7103262100\" target=\"_blank\">anti-human Fc single-domain antibody<\/a> <\/span>using a&nbsp;<a href=\"http:\/\/www.thermoscientific.com\/en\/product\/versette-automated-liquid-handler.html\" target=\"_blank\">&nbsp;Versette Automated Liquid Handler<\/a>&nbsp;&nbsp;(all from Thermo Scientific).&nbsp;During incubation, the micro-columns within each of the&nbsp;MSIA D.A.R.T.s selectively trapped&nbsp;adalimumab.<\/p>\n<p>The investigators<span class=\"thread\">&nbsp;used<\/span> a&nbsp;<a href=\"http:\/\/www.thermoscientific.com\/en\/product\/proswift-rp-protein-columns.html\" target=\"_blank\">ProSwift&nbsp;RP-4H LC column<\/a>&nbsp;for liquid chromatography and a<a href=\"http:\/\/www.thermoscientific.com\/en\/product\/q-exactive-hybrid-quadrupole-orbitrap-mass-spectrometer.html\" target=\"_blank\">&nbsp;Q Exactive hybrid quadrupole-Orbitrap mass spectrometer <\/a>with&nbsp;<a href=\"http:\/\/www.thermoscientific.com\/en\/product\/xcalibur-software.html\" target=\"_blank\">XCalibur software (revision 2.2)<\/a>&nbsp;(all from Thermo Scientific) for mass spectrometry analysis.<\/p>\n<p>LC-MS analysis generated four masses at<span class=\"thread\"> 148,080, 148,245, 148,407 and 148,565,<\/span> representing the addition of hexose groups to adalimumab. The masses at 148,203, 148,363 and 148,527 showed mass difference of ~123 Da, suggesting the presence of two sodium and two potassium adducts. The team also&nbsp;observed a linear dynamic range of 125 to 8,000 ng\/mL when intact adalimumab was detected with the native structure preserved. When&nbsp;the intact adalimumab was deglycosylated, they were able to detect&nbsp;adalimumab at concentrations as low as ~20 ng\/mL.<\/p>\n<p>Using&nbsp;MSIA, the investigators defined a more sensitive<span class=\"thread\"> system that enables researchers to obtain more<\/span> robust and reproducible results.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>References<\/strong><\/p>\n<p>1.&nbsp;Antwi, K.,&nbsp;Niederkofler, E.E. &amp;&nbsp;Kiernan, U.A. (2015) &#8220;A universal pre-clinical solution for the bioanalysis of fully human therapeutic monoclonal antibodies in plasma&#8221; [Poster]. Thermo Fisher.&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Monoclonal antibody (mAb) therapy&nbsp;is used to to stimulate the immune system to defend itself against particular target cells. As researchers explore mAb treatments further, they are increasingly using mass spectrometry to selectively analyze&nbsp;human therapeutic mAbs for therapeutic development. According to Antwi et al., current mass spectrometry methods to analyze mAbs are limited and result in<\/p>\n","protected":false},"author":7,"featured_media":5846,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_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":[12],"tags":[160],"division":[],"class_list":{"0":"post-5590","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-methods","8":"tag-mab-therapy","9":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- This site is optimized 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