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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

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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