Rapid loading of large sample volumes, analyte cleanup, and modified moving boundary transient isotachophoresis conditions for membrane preconcentration-capillary electrophoresis in small diameter capillaries

Andy J. Tomlinson, Linda M. Benson, Stephen Jameson, Stephen Naylor

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

Using a removable membrane preconcentration (mPC) cartridge, large sample volumes can be loaded prior to final assembly of the mPC capillary electrophoresis (CE) capillary. For narrow-bore (≤ 25 μm ID) uncoated mPC-CE capillaries, applied to peptide analysis, efficient moving boundary transient isotachphoresis (tITP) conditions at the onset of electrophoresis are described. The enhancement of mPC-CE-mass spectrometry (MS) technology afforded by rapid sample loading and modified moving boundary tITP conditions are demonstrated by analysis of major histocompatibility complex (MHC) class I peptides that were derived from a Kb precipitation of mouse EL-4 cells. Furthermore, we demonstrate the structural characterization of these immunologically significant molecules by mPC-CE-tandem mass spectrometry (mPC-CE-MS/MS).

Original languageEnglish (US)
Pages (from-to)1801-1807
Number of pages7
JournalELECTROPHORESIS
Volume17
Issue number12
DOIs
StatePublished - Jan 1 1996

Keywords

  • Capillary electrophoresis
  • Major histocompatibility complex class I peptides
  • Mass spectrometry
  • Preconcentration
  • Rapid loading

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