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CE coupled to MALDI with novel covalently coated capillaries
Stefan Bachmann1, Rainer Vallant, Rania Bakry
1Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens, University Innsbruck, Austria.
Electrophoresis
|February 18, 2010
Summary
New capillary coatings using polystyrene nanoparticles and fullerenes improve capillary electrophoresis (CE) for peptide and protein analysis. These coatings enhance reproducibility and stability, enabling effective separation and identification with mass spectrometry.
Area of Science:
- Analytical Chemistry
- Separation Science
- Biochemistry
Background:
- Capillary Electrophoresis (CE) offers flexibility for separating diverse compounds, including peptides and proteins.
- Adsorption of basic compounds, proteins, and peptides onto capillary walls necessitates surface modifications for reproducible CE separations.
- Capillary surface coatings are crucial for preventing adsorption and improving migration time and peak area repeatability.
Purpose of the Study:
- To investigate novel capillary coatings for enhanced CE analysis of peptides and protein digests.
- To evaluate the reproducibility, stability, and performance of derivatized polystyrene nanoparticle and fullerene coatings.
- To assess the combined potential of CE with MALDI-MS using these new coatings for complex sample analysis.
Main Methods:
- Development and application of new capillary coatings using derivatized polystyrene nanoparticles and fullerenes.
- Capillary electrophoresis (CE) separation of peptides and protein digests.
- Coupling of CE with Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-MS) for sample analysis.
- Database searching (Swiss-Prot) using MASCOT software for protein identification.
Main Results:
- Coated capillaries demonstrated excellent run-to-run (<0.5% RSD) and batch-to-batch (<9.5% RSD) reproducibility for migration times.
- The coatings exhibited high stability across a wide pH range (2.0-10.0).
- CE-MALDI-MS analysis of a five-protein digest mixture yielded sequence coverages between 36% and 68%.
Conclusions:
- Derivatized polystyrene nanoparticles and fullerenes provide effective capillary coatings for CE.
- These coatings significantly improve the reproducibility and stability of CE separations for peptides and proteins.
- The developed CE-MALDI-MS technology offers high separation efficiency and structure-selective identification capabilities for complex biological samples.
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