Capillary electrophoresis (CE) has emerged as one of the most promising methods for separation of proteins in their native states. Electrospray ionization allows transfer of proteins into the mass spectrometer while maintaining native conformations and non-covalent interactions between proteins. Integration of capillary electrophoresis with high performance mass spectrometers offers numerous opportunities for in-depth analysis of endogenous proteins, and we have focused on characterizing the proteins using ultraviolet photodissociation (UVPD), a high energy ion activation method that yields extensive fragmentation of even large proteins. UVPD causes extensive backbone cleavages while also allowing retention of post-translational modifications which facilitates their localization to specific residues. Here we describe the use of CE-UVPD-MS for characterization of biotherapeutics including antibody drug conjugates.

Capillary electrophoresis-mass spectrometry and ultraviolet photodissociation for characterization of proteins

ACS Fall 2021

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