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Successive Multiple Ionic-Polymer Layer Coatings for Intact Protein Analysis by Capillary Zone Electrophoresis–Mass Spectrometry: Application to Hemoglobin Analysis

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Capillary Electrophoresis-Mass Spectrometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2531))

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Abstract

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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Author information

Author notes

  1. Liesa Salzer

    Present address: Analytical BioGeoChemistry (BGC), Helmholtz Zentrum München, Neuherberg, Germany

Authors and Affiliations

  1. Faculty of Chemistry, Aalen University, Aalen, Germany

    Liesa Salzer, Alexander Stolz, Alisa Höchsmann & Christian Neusüß

  2. Institut des Biomolécules Max Mousseron, UMR CNRS, Université de Montpellier, Montpellier, France

    Laura Dhellemmes, Laurent Leclercq & Hervé Cottet

Authors
  1. Liesa Salzer
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  2. Alexander Stolz
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  3. Laura Dhellemmes
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  4. Alisa Höchsmann
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  5. Laurent Leclercq
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  6. Hervé Cottet
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  7. Christian Neusüß
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Corresponding author

Correspondence to Christian Neusüß .

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Editors and Affiliations

  1. Aalen University, Aalen, Baden-Württemberg, Germany

    Christian Neusüß

  2. Aalen University, Aalen, Baden-Württemberg, Germany

    Kevin Jooß

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Salzer, L. et al. (2022). Successive Multiple Ionic-Polymer Layer Coatings for Intact Protein Analysis by Capillary Zone Electrophoresis–Mass Spectrometry: Application to Hemoglobin Analysis. In: Neusüß, C., Jooß, K. (eds) Capillary Electrophoresis-Mass Spectrometry . Methods in Molecular Biology, vol 2531. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2493-7_5

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  • DOI: https://doi.org/10.1007/978-1-0716-2493-7_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2492-0

  • Online ISBN: 978-1-0716-2493-7

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