Coupling ion mobility separations, collisional activation techniques, and multiple stages of MS for analysis of complex peptide mixtures.
An ion trap/ion mobility/quadrupole/collision cell/time-of-flight mass spectrometer that incorporates a differentially pumped orifice-skimmer cone region at the back of the drift tube has been developed for the analysis of peptide mixtures. The combined approach allows a variety of strategies to be employed for collisionally activating ions, and fragments can be monitored by subsequent stages of mass spectrometry in a parallel fashion, as described previously (Anal. Chem. 2000, 72, 2737). Here, we describe the overall experimental approach in detail. Applications involving different aspects of the initial mobility separation and various collisional activation and parallel sequencing strategies are illustrated by examining several simple peptide mixtures and a mixture of tryptic peptides from beta-casein. Detection limits associated with various experimental configurations and the utility for analysis of complex systems are discussed.
Hoaglund-Hyzer
Cherokee S
CS
Department of Chemistry, Indiana University, Bloomington 47405, USA.
Lee
Young Jin
YJ
Counterman
Anne E
AE
Clemmer
David E
DE
D000465:Algorithms; D000595:Amino Acid Sequence; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D011492:Protein Hydrolysates / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
D007202:Indicators and Reagents; D010455:Peptides; D011492:Protein Hydrolysates