Recent developments and applications of capillary and microchip electrophoresis in proteomics and peptidomics (mid‐2018–2022)

Abstract

This review gives a wide overview of recent advances and applications of capillary electrophoresis and microchip capillary electrophoresis methods in the fields of proteomics and peptidomics in the period from mid‐2018 up to the end of 2022. The methodological topics covering sample preparation and concentration techniques, hyphenation of capillary electrophoresis methods with mass spectrometry, and multidimensional separations by on‐line or off‐line coupled different capillary electrophoresis and liquid chromatography techniques are described and new developments in both bottom‐up and top‐down approaches in proteomics are presented. In addition, various applications of capillary electrophoresis methods in proteomic and peptidomic studies are demonstrated. They include monitoring of protein posttranslational modifications and applications in biological and biochemical research, clinical peptidomics and proteomics, and food analysis.

Recent developments in capillary and microchip electroseparations of peptides (2021–mid‐2023)

“…The important role of multidimensional separations of biopeptides, peptide fragments of proteins and proteins in proteomic, peptidomic, and metabolomic studies was highlighted in the recent reviews [13, 39, 41]. Two‐ or multidimensional peptide separations are based on the combination of two or more separation methods with different separation principles, for example, LC and electrophoresis [181, 182].…”

“…In these cases, a combination of two or more complementary separation principles, generating 2D or multidimensional separation systems, such as IEF and SDS-PAGE in the classical 2D gel electrophore-sis (2D-GE) [178,179] or in the differential or comparative 2D-GE [180] is needed. The important role of multidimensional separations of biopeptides, peptide fragments of proteins and proteins in proteomic, peptidomic, and metabolomic studies was highlighted in the recent reviews [13,39,41]. Two-or multidimensional peptide separations are based on the combination of two or more separation methods with different separation principles, for example, LC and electrophoresis [181,182].…”

“…These-omics technologies-are complex and interconnected disciplines. Both, structure and function of proteins are often investigated by means of their enzymatically generated peptide fragments in the so called bottom-up or shotgun proteomics [10][11][12][13]. These peptide fragments are separated and identified by liquid chromatography (LC), 2-DE, and capillary electrophoresis (CE) methods online coupled with MS detection [14][15][16][17][18][19].…”

“…In addition, for understanding of both normal and pathological physiological processes, a comprehensive investigation of the whole peptide set (peptidome) of a cell, organ, or organism at given time frame is necessary as well. This is the subject of peptidomics-a bridge between proteomics and metabolomics [12,13,[20][21][22][23]. Thus, separation, analysis, isolation, purification, and characterization of peptides by capillary and microchip electromigration methods are one of the most challenging tasks of these high-performance separation techniques.…”

Investigating the Merits of Microfluidic Capillary Zone Electrophoresis–Mass Spectrometry (CZE-MS) in the Bottom-Up Characterization of Larger RNAs

“…The compact design necessary to fit the entire separation channel onto a relatively small chip translates into greater drops of potential per length unit, which can lead to shorter analysis time with no significant deterioration of electrophoretic resolution. The capabilities of microfluidic CZE-MS in the characterization of sample mixtures have been successfully demonstrated by the analysis of intact antibodies, , protein digests, − metabolite extracts, − and intact oligonucleotides . In this report, we evaluated the merits of the technique in the characterization of typical digestion mixtures encountered during the bottom-up analysis of NA substrates.…”