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Electrifying the Molecules of Life : Peptide and Protein Analysis by Capillary Electrophoresis Coupled to Electrospray Ionization Mass Spectrometry
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- Wetterhall, Magnus, 1973- (författare)
- Uppsala universitet,Kemiska institutionen
- Bergquist, Jonas (preses)
- Markides, Karin E. (preses)
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- ISBN 9155459471
- Uppsala : Acta Universitatis Upsaliensis, 2004
- Engelska 44 s.
- Serie: Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, 1104-232X ; 969
- Doktorsavhandling (övrigt vetenskapligt/konstnärligt)
Abstract
Ämnesord
Stäng
- This thesis describes the current status and novel aspects of the analysis of the molecules of life, i.e. peptides and proteins, using capillary electrophoresis (CE) coupled to mass spectrometry (MS) via (sheathless) electrospray ionization (ESI). Early reports of sheathless CE-ESI-MS were plagued by limited lifetimes of the electrospray emitter. In this thesis, two new approaches, the Black Dust and the Black Jack methods, utilizing polymer-embedded graphite instead of noble metals are presented. These emitters have shown improved long-term stability and proven excellent for sheathless electrospray operation. Failure of an emitter is often caused by electrochemical reactions occurring at the emitter-liquid interface. The electrochemical properties of the graphite coated emitters were therefore evaluated by classical electrochemical methods, such as cyclic voltammetry and chronoamperometry. The graphite coated emitters showed excellent electrochemical stability and properties compared to noble metal and polymer configurations. Analyte-wall interactions have long been known to cause problems in the CE analysis of biomolecules. This can be circumvented by internal modification of the capillary walls. Additionally, it is of outermost importance to have a stable and sufficiently high electroosmotic flow (EOF) to sustain the electrospray, when using a sheathless approach. New monomer and polymer coatings are presented for rapid and high-efficient CE-ESI-MS separations of peptides and proteins. Furthermore, the use of CE-ESI coupled to Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) shows great potential for rapid proteomic probing of human cerebrospinal fluid. The results are comparable with more established techniques, such as liquid chromatography and two-dimensional gel electrophoresis coupled to MS. However, the CE-ESI-FTICRMS analysis has significantly lower sample consumption and faster analysis time compared to the other techniques. The applications and use of CE-ESI-MS is expected to have a bright future with continued growth as current trends of multidimensional hyphenation and microfabricated devices are further developed and explored.
Ämnesord
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Nyckelord
- Analytical chemistry
- Capillary electrophoresis (CE)
- Electrospray ionization (ESI)
- Mass spectrometry (MS)
- Peptides
- Proteins
- Cerebrospinal fluid (CSF)
- Graphite coating
- Monomer coating
- Polymer coating
- Analytisk kemi
- Analytical chemistry
- Analytisk kemi
Publikations- och innehållstyp
- vet (ämneskategori)
- dok (ämneskategori)
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