Using accurate mass electrospray ionization-time-of-flight mass spectrometry with in-source collision-induced dissociation to sequence peptide mixtures.
Although data-dependent LC-MS-MS with database searching has become au courant for identifying proteins, the technique is constrained by duty-cycle inefficiency and the inability of most tandem mass analyzers to accurately measure peptide product ion masses. In this work, a novel approach is presented for simultaneous peptide fragmentation and accurate mass measurement using in-source collision-induced dissociation (CID) on electrospray ionization (ESI)-time-of-flight (TOF) MS. By employing internal mass reference compounds, mass measurement accuracy within +/-5 ppm for tryptic peptide precursors and +/-10 ppm for most sequence-specific product ions was consistently achieved. Analysis of a complex solution containing several digested protein standards did not adversely affect instrument performance.
Williams
Jon D
JD
Discovery Research, GlaxoSmithKline, 5 Moore Drive, Research Triangle Park, NC 27709-3398, USA. Jon.D.Williams@gsk.com
Flanagan
Michael
M
Lopez
Linda
L
Fischer
Steve
S
Miller
Luke A D
LA
D002851:Chromatography, High Pressure Liquid; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014357:Trypsin / Q000737:chemistry
D010455:Peptides; D014357:Trypsin