Advanced online mass spectrometry detection of proteins separated by capillary isoelectric focusing after sequential injection

Csilla Páger, Nikoleta Biherczová, Roland Ligetvári, Balázs Viktor Berkics, Tamás Pongrácz, Viktor Sándor, Anita Bufa, Viktória Poór, Andrea Vojs Staňová, F. Kilár

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1 Citation (Scopus)

Abstract

Capillary isoelectric focusing hyphenated with mass spectrometry detection, following the sequential injection of the carrier ampholytes and the sample zone, is highly efficient for the characterization of proteins. The main advantage of the sequential injection protocol is that ampholytes, with pH ranges, which are not supposed to cover the isoelectric points of the sample components, can be used for separation. The method then allows online mass spectrometry detection of separated analytes either in the absence (substances that have left the pH gradient) or in the presence of low-level ampholytes (substances that are migrating within the pH gradient). The appearance of the substances within, or outside the pH gradient depends on, e.g., the composition of the ampholytes (broad or narrow pH range) or on the composition of electrolyte solutions. The experiments performed in coated capillaries (with polyvinyl alcohol or with polyacrylamide) show that the amount and the injection length of the ampholytes influence the length of the pH gradient formed in the capillary.

Original languageEnglish
Pages (from-to)4825-4834
Number of pages10
JournalJournal of Separation Science
Volume40
Issue number24
DOIs
Publication statusPublished - Dec 1 2017

Keywords

  • capillary isoelectric focusing
  • carrier ampholytes
  • mass spectrometry
  • proteins
  • sequential injection

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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    Páger, C., Biherczová, N., Ligetvári, R., Berkics, B. V., Pongrácz, T., Sándor, V., Bufa, A., Poór, V., Vojs Staňová, A., & Kilár, F. (2017). Advanced online mass spectrometry detection of proteins separated by capillary isoelectric focusing after sequential injection. Journal of Separation Science, 40(24), 4825-4834. https://doi.org/10.1002/jssc.201700695