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Capillary electrophoresis-off-line-matrix-assisted laser desorption/ionisation mass spectrometry of proteins and peptides using cationic-coated capillaries
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry.
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Manuscript (Other academic)
URN: urn:nbn:se:uu:diva-91604OAI: oai:DiVA.org:uu-91604DiVA: diva2:164396
Available from: 2004-04-23 Created: 2004-04-23 Last updated: 2011-03-21
In thesis
1. Tailormade Surfaces for Extended CE Applications
Open this publication in new window or tab >>Tailormade Surfaces for Extended CE Applications
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

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.

In the pharmaceutical industry, favorable pharmacokinetic properties of a candidate drug, such as high bioavailability after oral administration, are crucial for a high success rate in clinical development. Tools for prediction of biopharmaceutically relevant drug properties are important in order to identify and discard poor candidate drugs as soon as possible. In this thesis, a membrane mimetic coating was developed by electrostatically immobilizing liposomes to the capillary wall, via an anchoring sublayer of Q-agarose. The liposome-coated capillaries were demonstrated in on-line CE-MS for prediction of drug membrane permeability.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2004. 50 p.
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 962
Analytical chemistry, capillary electrophoresis, mass spectrometry, surface properties, coatings, polymers, liposomes, proteins, drugs, Analytisk kemi
National Category
Analytical Chemistry
urn:nbn:se:uu:diva-4217 (URN)91-554-5934-X (ISBN)
Public defence
2004-05-14, B42, BMC, Husargatan 3, Uppsala, 10:15
Available from: 2004-04-23 Created: 2004-04-23 Last updated: 2012-02-08Bibliographically approved

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Bergquist, Jonas
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