Top-Down Characterization of an Intact Monoclonal Antibody Using Activated Ion Electron Transfer Dissociation.
Top-Down Characterization of an Intact Monoclonal Antibody Using Activated Ion Electron Transfer Dissociation.
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DOI：
10.1021/acs.analchem.0c00705
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发表时间：
2020-08-04
影响因子：
7.4
通讯作者：
Coon JJ
中科院分区：
文献类型：
--
作者：
Lodge JM;Schauer KL;Brademan DR;Riley NM;Shishkova E;Westphall MS;Coon JJ
Monoclonal antibodies (mAbs) are important therapeutic glycoproteins, but their large size and structural complexity make them difficult to rapidly characterize. Top-down mass spectrometry (MS) has potential to overcome challenges of other common approaches by minimizing sample preparation and preserving endogenous modifications. However, comprehensive mAb characterization requires generation of many, well-resolved fragments and remains challenging. While ETD retains modifications and cleaves disulfide bonds – making it attractive for mAb characterization – it can be less effective for precursors having high m/z values. Activated ion electron transfer dissociation (AI-ETD) uses concurrent infrared photoactivation to promote product ion generation and has proven effective in increasing sequence coverage of intact proteins. Here we present the first application of AI-ETD to mAb sequencing. For the standard NIST mAb we observe a high degree of complementarity between fragments generated using standard ETD with a short reaction time and AI-ETD with a long reaction time. Most importantly, AI-ETD reveals disulfide-bound regions that have been intractable, thus far, for sequencing with top-down MS. We conclude AI-ETD has the potential to rapidly and comprehensively analyze intact mAbs.
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影响因子：
7.4
作者：
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通讯作者：
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DOI：
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通讯作者：
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