CE‐MS for the analysis of intact proteins 2010–2012

Abstract

Since its introduction in 1987, CE-MS has become an increasingly important technique for the analysis of biomolecules. Since our previous update on CE-MS methods within the field of intact protein analysis (Electrophoresis 2011, 32, 66-82), a variety of interesting methodological improvements and applications have been reported in literature. Therefore, this article presents an overview of the development and application of CE-MS for intact protein analysis as published between June 2010 and June 2012. The article is divided in sections that treat CE coupled to MS through ESI, MALDI, and ICP ionization, respectively. In the section about CE-ESI-MS, technological developments with respect to CE-MS interfacing, prevention of protein adsorption, and chip-based CE-MS are treated in more detail. Novel interfacing strategies and the development of improved capillary coating strategies appeared to be the major developments. Furthermore, in all sections, the applicability of CE-MS for intact protein analysis is demonstrated by representative examples, including important developments in the fields of biopharmaceutical characterization and the analysis of proteins in biological samples. Finally, some general conclusions and future perspectives are given.

Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis

“…This technique can provide important information on identity, glycoforms, degradation, and impurities of protein therapeutics [86, 87]. It is possible to couple CE to MS using different ionization techniques, as has been described in several excellent reviews [88, 89]. For this review, only the recent advances regarding the development and application of the ESI interfaces will be highlighted.…”

Mass spectrometry for glycosylation analysis of biopharmaceuticals

“…CZE-MS has been most extensively used for the characterization of intact glycoproteins [69]. Most efficient separations of glycoproteins are obtained when using low-pH background electrolytes in combination with coatings that minimize or cancel out electroosmotic flow.…”

Developments in Interfacing Designs for CE–MS: Towards Enabling Tools for Proteomics and Metabolomics

“…top-down proteomics), as well as on the peptide level (i.e. bottom-up proteomics) [3,53,54]. In this section, the potential of CE-MS using novel interfacing techniques is demonstrated for some illustrative proteomics studies.…”

“…In this section, the potential of CE-MS using novel interfacing techniques is demonstrated for some illustrative proteomics studies. For a comprehensive overview of CE-MS-based proteomics studies, we refer to the recent reviews of Haselberg et al [3], Heemskerk et al [53] and Zhao et al [2].…”

“…Capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) has emerged as a powerful tool for the analysis of charged compounds in biological samples [1][2][3]. CE-MS can be used for the characterization of a wide variety of analytes, e.g., metabolites, peptides, glycans and intact proteins [4][5][6].…”