Tailormade Surfaces for Extended CE Applications

The combination of capillary electrophoresis (CE) and mass spectrometry (MS) constitutes a powerful microanalytical system in the fields of biology, medicine and chemistry. This thesis describes the development of three novel capillary coatings and demonstrates how these extend the utility of CE as a high-efficiency separation technique in protein analysis and biopharmaceutical drug screening. Due to the rapidly growing interest in characterizing the human proteome, there is an increased need for rapid protein separations. The use of CE in protein analysis is, however, nontrivial due to problems with protein adsorption to the fused-silica capillary walls. In this thesis, this problem was addressed by developing two novel, physically adsorbed, cationic polymer surface coatings, denoted PolyE-323 and Q-agarose. By using simple rinsing protocols, highly reproducible coatings, stable over a wide range of pH 2-11 were generated. Successful protein separations using cationic-coated capillaries in CE-MS, equipped with either electrospray ionization (ESI) or matrix-assisted laser desorption/ionization (MALDI), has been demonstrated…


1 Introduction
2 Capillary electrophoresis
2.1 Basic concepts of CE
2.2 CE coupled to mass spectrometry
3 Capillary coatings
3.1 Deactivating coatings
3.1.1 Neutral deactivation layers
3.1.2 Cationic deactivation layers
3.2 Intermediate-layer coatings
3.3 Selective coatings
4 Protein analysis
4.1 Analyte-wall interactions
4.2 CE-analysis of proteins in biofluids
4.3 Mass spectrometric detection of proteins
5 Biopharmaceutical drug screening
5.1 Drug absorption
5.2 Drug screening
6 Conclusions
7 Acknowledgements
8 Summary in Swedish
9 References

Author: Ullsten, Sara

Source: Uppsala University Library

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