LC/MS at the whole protein level: Studies of biomolecular structure and interactions using native LC/MS and cross-path reactive chromatography (XP-RC) MS

Review article, 2018

Interfacing liquid chromatography (LC) with electrospray ionization (ESI) to enable on-line MS detection had
been initially implemented using reversed phase LC, which in the past three decades remained the default type of
chromatography used for LC/MS and LC/MS/MS studies of protein structure. In contrast, the advantages of other
types of LC as front-ends for ESI MS, particularly those that allow biopolymer higher order structure to be
preserved throughout the separation process, enjoyed relatively little appreciation until recently. However, the
past few years witnessed a dramatic surge of interest in the so-called “native” (with “non-denaturing” being
perhaps a more appropriate adjective) LC/MS and LC/MS/MS analyses within the bioanalytical and biophysical
communities. This review focuses on recent advances in this field, with an emphasis on size exclusion and ion
exchange chromatography as front-end platforms for protein characterization by LC/MS. Also discussed are the
benefits provided by the integration of chemical reactions in the native LC/MS analyses, including both ion
chemistry in the gas phase (e.g., limited charge reduction for characterization of highly heterogeneous biopolymers)
and solution-phase reactions (using the recently introduced technique cross-path reactive chromatography).