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Complete Characterization of a Cysteine-linked Antibody-Drug Conjugate Performed on a Hybrid Quadrupole-Orbitrap Mass Spectrometer with High Mass Range

Orbitrap_SciLib
Reputable Mentor II
Reputable Mentor II
Bailey A, Damoc E, Houel S, Scheffler K, Josephs J.
ASMS 2016 Poster
We have modified the instrument control software of a benchtop quadrupole-Orbitrap mass spectrometer to add native MS capability. In this study we demonstrate complete characterization of Brentuximab vedotin, a cysteine-linked ADC, which requires native MS conditions for intact analysis. We demonstrate preservation of non-covalent bonding of antibody subunits during electrospray ionization. HMR mode can be turned off for peptide mapping. We use trypsin peptide mapping approach with HCD fragmentation to achieve 99% coverage of the Brentuximab vedotin sequence using a single LC-MS analysis of a 90 min reverse phase gradient. Finally, we demonstrate that signature ions specific for HCD fragmentation of Brentuximab vedotin can be utilized to increase MS/MS assignment confidence.


Thermo Fisher Scientific
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Last update:
‎10-15-2021 11:44 AM
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