Capillary liquid chromatography/atmospheric-pressure matrix-assisted laser desorption/ionisation ion trap mass spectrometry: a comparison with liquid chromatography/matrix-assisted laser desorption/ionisation time-of-flight and liquid chromatography/electrospray ionisation quadrupole time-of-flight for the identification of tryptic peptides.
The atmospheric-pressure matrix-assisted laser desorption/ionisation quadrupole ion trap (AP-MALDI-QIT) analysis of tryptic peptides is reported following capillary liquid chromatographic (LC) separation and direct analysis of a protein digest. Peptide fragments were identified by peptide mass fingerprinting from mass spectrometric data and sequence analysis obtained by tandem mass spectrometry of the principal mass spectral peaks using a data-dependent scanning protocol. These data were compared with those from mass spectrometric analysis using capillary LC/MALDI-time-of-flight (TOF) and capillary LC/electrospray ionisation (ESI)-quadrupole TOF. For all three configurations the resulting data were searched against the MSDB database, using MASCOT and the sequence coverage compared for each technique. Complementary data were obtained using the three techniques.
Creaser
Colin S
CS
School of Biomedical and Natural Sciences, Nottingham Trent University, Nottingham NG11 8NS, UK. colin.creaser@ntu.ac.uk
Green
Philip S
PS
Kilby
Peter M
PM
Ratcliffe
Lucy
L
D000595:Amino Acid Sequence; D000818:Animals; D001274:Atmospheric Pressure; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification*; D010449:Peptide Mapping; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D010446:Peptide Fragments; D012710:Serum Albumin, Bovine