Rapid loading of large sample volumes, analyte cleanup, and modified moving boundary transient isotachophoresis conditions for membrane preconcentration-capillary electrophoresis in small diameter capillaries.
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 (< or = 25 microns 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).
Tomlinson
A J
AJ
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA.
Benson
L M
LM
Jameson
S
S
Naylor
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000737:chemistry; D013058:Mass Spectrometry; D051379:Mice; D010455:Peptides / Q000032:analysis / Q000737:chemistry
D015395:Histocompatibility Antigens Class I; D010455:Peptides