Efficient and highly reproducible capillary electrophoresis-mass spectrometry of peptides using Polybrene-poly(vinyl sulfonate)-coated capillaries

J.R. Catai, Javier Sastre Torano, GJ de Jong, GW Somsen*

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    The potential of capillaries noncovalently coated with a bilayer of oppositely charged polymers for the analysis of peptides by CE-MS was investigated. Bilayer coatings were produced by subsequently rinsing fused-silica capillaries with a solution of Polybrene (PB) and poly(vinyl sulfonate) (PVS). The PB-PVS coating showed to be fully compatible with MS detection causing no ionization suppression or background signals. The bilayer coating provided a considerable EOF at low pH, thereby facilitating the fast separation of peptides using a BGE of formic acid (pH 2.5). Under optimized CE-MS conditions, for enkephalin peptides high separation efficiencies were obtained with plate numbers in the range of 300 000-500 000. It is demonstrated that both the cancellation of the hydrodynamic capillary flow induced by the nebulizer gas and a sufficiently high-data acquisition rate are crucial for achieving these efficiencies. The overall performance of the CE-MS system using PB-PVS-coated capillaries was evaluated by the analysis of a tryptic digest of cytochrome c. The system provided an efficient separation of the peptide mixture, which could be effectively monitored by MS/MS detection allowing identification of at least 13 peptides within a time interval of 1.5 min. In addition, the PB-PVS coating proved to be very consistent yielding stable CE-MS patterns with highly favorable migration time reproducibilities (RSDs <1% over a 3-day period).

    Original languageEnglish
    Pages (from-to)2091-2099
    Number of pages9
    JournalElectrophoresis
    Volume27
    Issue number11
    DOIs
    Publication statusPublished - Jun 2006

    Keywords

    • capillary electrophoresis
    • electrospray ionization
    • mass spectrometry
    • noncovalent bilayer coatings
    • peptides
    • BILAYER-COATED CAPILLARIES
    • ELECTROSPRAY-IONIZATION
    • BASIC-PROTEINS
    • SEPARATION
    • COATINGS
    • CHROMATOGRAPHY
    • ADSORPTION

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