Native Capillary Electrophoresis-Mass Spectrometry for Protein analysis:
Current Progress and Perspectives
Abstract
Native mass spectrometry is a rapidly emerging technique for fast and
sensitive structural analysis of protein constructs, maintaining the
protein higher order structure. The coupling with electromigrative
separation techniques under native conditions enables the
characterization of proteoforms and highly complex protein mixtures. In
this review, we present an overview of current native CE-MS technology.
First, the status of native separation conditions is described for
capillary zone electrophoresis (CZE), affinity capillary electrophoresis
(ACE), and capillary isoelectric focusing (CIEF), as well as their
chip-based formats, including essential parameters such as electrolyte
composition and capillary coatings. Further, conditions required for
native ESI-MS of (large) protein constructs, including instrumental
parameters of QTOF and Orbitrap systems, as well as requirements for
native CE-MS interfacing are presented. On this basis, methods and
applications of the different modes of native CE-MS are summarized and
discussed in the context of biological, medical, and biopharmaceutical
questions. Finally, key achievements are highlighted and concluded,
while remaining challenges are pointed out.