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Advanced online mass spectrometry detection of proteins separated by capillary isoelectric focusing after sequential injection
C. Páger, N. Biherczová, R. Ligetvári, BV. Berkics, T. Pongrácz, V. Sándor, A. Bufa, V. Poór, A. Vojs Staňová, F. Kilár,
Jazyk angličtina Země Německo
Typ dokumentu časopisecké články
PubMed
29064636
DOI
10.1002/jssc.201700695
Knihovny.cz E-zdroje
- MeSH
- amfolytové směsi MeSH
- elektroforéza kapilární * MeSH
- hmotnostní spektrometrie * MeSH
- isoelektrická fokusace * MeSH
- koncentrace vodíkových iontů MeSH
- proteiny izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
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.
Citace poskytuje Crossref.org
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