Characterization of the N-glycosylation Pattern of Antibodies by ESI – and MALDI Mass Spectrometry
| Type: | Application |
Application Note |
Expression Proteomics, Gen. Pharma & Chemistry |
| Number: | Technology |
ET-17 / MT-99 |
MALDI-TOF, UHR-TOF |
| Year | Products |
2010 |
maXis, ultrafleXtreme, EASY-nano LC, PROTEINEER fc II, BioTools |
| Author | |
Arndt Asperger, Dirk Wunderlich, Marcus Macht, Pierre-Olivier Schmit; Bruker Daltonik, Bremen, Germany |
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| Reference | |
Bruker Daltonics Application Note ET-17 / MT-99 |
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Abstract |
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Analysis of the N-glycosylation pattern on antibodies is described using complementary mass spectrometric strategies based on both, top-down ESI-UHR-TOF and bottom-up LC-MALDI-TOF/TOF. Fast LC-ESI-UHR-TOF analysis, performed on the Bruker maXis™, provides high-resolution, high-mass accuracy (confident low ppm) data for both, intact antibodies and released antibody heavy chains allowing a rapid assignment of the major N-glycosylation isoforms. Bruker´s new ultrafleXtreme™ platform facilitates in-depth characterization of the antibodies´ N-glycosylation patterns via LC-MALDI-TOF/TOF analysis of the N-glycopeptides generated from digested antibodies. The improved resolution provided by the instrument delivers an extremely detailed picture of the highly complex patterns of N-linked glycans present on antibodies from different origins. As a unique feature, MALDI-TOF/TOF data generated from N-glycopeptides, simultaneously provide information in the same spectrum including both the peptide sequence and the structure of the N-linked glycan moiety. Selective screening and subsequent analysis of spectra of N-glycopeptides from large LC-MALDI-MS/MS datasets is supported by dedicated features implemented in Bruker´s latest software tools. |
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Related Products |
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ultrafleXtreme series |
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