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Open Access Green as soon as Postprint is submitted to ZB.
Successive multiple ionic-polymer layer coatings for intact protein analysis by capillary zone electrophoresis-mass spectrometry: Application to hemoglobin analysis.
Methods Mol. Biol. 2531, 69-76 (2022)
Adsorption of analytes, e.g., proteins, often interfere with separation in CE, due to the relatively large surface of the narrow capillary. Coatings often are applied to prevent adsorption and to determine the electroosmotic flow (EOF), which is of major importance for the separation in CE. Successive multiple ionic-polymer layer (SMIL) coatings are frequently used for protein analysis in capillary electrophoresis resulting in high separation efficiency and repeatability. Here, the coating procedure of a five-layer SMIL coating is described using quaternized diethylaminoethyl dextran (DEAEDq) as polycation and poly(methacrylic acid) (PMA) as polyanion. Depending on the analyte, different polyions may be used to increase separation efficiency. However, the coating procedure remains the same.To demonstrate the applicability of SMIL coatings in CE-MS, human hemoglobin was measured in a BGE containing 2 M acetic acid. DEAEDq-PMA coating was found to be the most suitable for hemoglobin analysis due to relatively low reversed electroosmotic mobility leading to increased electrophoretic resolution of closely related proteoforms. Thereby, not only alpha and beta subunit of the hemoglobin could be separated, but also positional isoforms of glycated and carbamylated species were separated within 24 min.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Ce-ms ; Cze ; Hemoglobin ; Polyelectrolytes ; Smil Coatings ; Static Adsorbed Capillary Coatings
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Book Volume Title
Capillary Electrophoresis-Mass Spectrometry
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 2531,
Pages: 69-76
Publisher
Springer
Publishing Place
Berlin [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Research Unit Analytical BioGeoChemistry (BGC)