Institute of Physical and Theoretical Chemistry

New coating materials for CE-SDS, CE-IEF and CE-MS coupling

The use of capillary coatings to suppress analyte adsorption onto the capillary surface is mandatory for the analysis of proteins, peptides, but also other types of analytes such as biogenic amines. Additionally, the capillary coatings show a significant impact on analyte resolution and separation via the induced electroosmotic flow. We investigate the use of physically adsorbed coating materials on the one hand and covalently coupled coating materials on the other hand. The physically adsorbed coatings can easily be attached to the capillary surface by fast and simple coating procedures. They allow robust separations with high repeatability and they are sufficiently stable for the direct coupling with mass spectrometric detection.

We are engaged in the development and characterization of cationic coating materials, especially for the CE-MS analysis of peptides and intact proteins. Our results for the separation of intact proteins and glycopeptide isoforms with self-synthesized cationic capillary coatings show that the induced moderate electroosmotic flow provides excellent separation characteristics which are not accessible with comparable commercial coating materials. At the same time, the self-synthesized coatings are impressively stable. This also accounts for covalently coupled coating materials which provide an even better suppression of protein adsorption. Here, we focus on the development of simple, but at the same time robust coating procedures, especially for the analysis of intact proteins.

Publications:

  1. Statically Adsorbed Coatings for High Separation Efficiency and Resolution in CE-MS Peptide Analysis: Strategies and Implementation.
    M. Pattky, K. Barkovits, O. Weiergräber, C. Huhn, Meth. Mol. Biol. 2016, 1483, 53-75

  2. Spermidine, but not spermine, is essential for pigment pattern formation in zebrafish
    H. G.     Frohnhöfer, S. Geiger-Rudolph, M. Pattky, M. Meixner, C. Huhn, H. M. Maischein, R. Geisler, I. Gehring, F. Maderspacher, C. Nüsslein-Volhard, U. Irion, Biol. Open. 2016, 5, 736-744.

  3. Structure characterization of unexpected covalent O-sulfonation and ion-pairing on an extremely hydrophilic peptide with CE-MS and FT-ICR-MS
    M. Pattky, S. Nicolardi, B. Santiago-Schübel, D. Sydes, Y. E. van der Burgt, A. N. Klein, J. Mohrlüder, K. Hänel, J. Kutzsche, S. A. Funke, D. Willbold, S. Willbold, C. Huhn, Anal. Bioanal. Chem. 2015, 407, 6637-6655

  4. Statically Adsorbed Coatings for High Separation Efficiency and Resolution in CE-MS Peptide Analysis: Strategies and Implementation.
    M. Pattky, K. Barkovits, O. Weiergräber, C. Huhn, Meth. Mol. Biol. 2016, 1483, 53-75

  5. Advantages and limitations of a new cationic coating inducing a slow electroosmotic flow for CE-MS peptide analysis: a comparative study with commercial coatings
    M. Pattky, C. Huhn*, Anal. Bioanal. Chem. 2013, 405, 225-237

  6. Relevance and use of capillary coatings in capillary electrophoresis-mass spectrometry
    C. Huhn, R. Ramautar, M. Wuhrer, G. W. Somsen, Anal. Bioanal. Chem. 2010, 396, 297-314