Abstract
Capillary electrophoresis (CE) played an important role in developments in the life sciences. The technique is nowadays used for the analysis of both large and small molecules in applications where it performs better than or is complementary to liquid chromatographic techniques. In this review, principles of different electromigration techniques, especially capillary isoelectric focusing (CIEF), capillary gel (CGE) and capillary zone electrophoresis (CZE), are described and recent developments in instrumentation, with an emphasis on mass spectrometry (MS) coupling and microchip CE, are discussed. The role of CE in the life sciences is shown with applications in which it had a high impact over the past few decades. In this context, current practice for the characterization of biopharmaceuticals (therapeutic proteins) is shown with CIEF, CGE and CZE using different detection techniques, including MS. Subsequently, the application of CGE and CZE, in combination with laser induced fluorescence detection and CZE-MS are demonstrated for the analysis of protein-released glycans in the characterization of biopharmaceuticals and glycan biomarker discovery in biological samples. Special attention is paid to developments in capillary coatings and derivatization strategies for glycans. Finally, routine CE analysis in clinical chemistry and latest developments in metabolomics approaches for the profiling of small molecules in biological samples are discussed. The large number of CE applications published for these topics in recent years clearly demonstrates the established role of CE in life sciences.
Highlights
After a long development, capillary electromigration techniques are very strong in various fields [1]
capillary electrophoresis (CE) became strong in life sciences with the rapid and efficient analysis of small highly polar compounds and large biomolecules such as deoxyribonucleic acid (DNA), peptides and proteins
The analysis of released glycans in contrast is usually performed with capillary zone electrophoresis (CZE)- or capillary gel electrophoresis (CGE) coupled to laser-induced fluorescence (LIF) detection, because detection limits with CE-mass spectrometry (MS) are not sufficient due to the low injection volumes in CE
Summary
Capillary electromigration techniques are very strong in various fields [1]. In 1937 Tiselius described protein separations using an U-tube [3] but only a separation of two components was obtained and the applicability was still limited. A main contribution was offered by Martin who obtained an efficient separation of chloride, acetate, aspartate and glutamate using displacement electrophoresis (isotachophoresis, ITP) [4]. CE is very suitable for the separation of polar and ionogenic compounds and the applicability in main fields of the life sciences is shown. In clinical chemistry and metabolomics various applications demonstrate the possibilities of capillary electromigration separation techniques for the analysis of complex samples, especially for profiling and biomarker discovery. Advanced systems and perspectives of CE are discussed and some figures illustrate that CE is a strong technique in life sciences
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