Toward plasma proteome profiling with ion mobility-mass spectrometry.
Differential, functional, and mapping proteomic analyses of complex biological mixtures suffer from a lack of component resolution. Here we describe the application of ion mobility-mass spectrometry (IMS-MS) to this problem. With this approach, components that are separated by liquid chromatography are dispersed based on differences in their mobilities through a buffer gas prior to being analyzed by MS. The inclusion of the gas-phase dispersion provides more than an order of magnitude enhancement in component resolution at no cost to data acquisition time. Additionally, the mobility separation often removes high-abundance species from spectral regions containing low-abundance species, effectively increasing measurement sensitivity and dynamic range. Finally, collision-induced dissociation of all ions can be recorded in a single experimental sequence while conventional MS methods sequentially select precursors. The approach is demonstrated in a single, rapid (3.3 h) analysis of a plasma digest sample where abundant proteins have not been removed. Protein database searches have yielded 731 high confidence peptide assignments corresponding to 438 unique proteins. Results have been compiled into an initial analytical map to be used -after further augmentation and refinement- for comparative plasma profiling studies.
Valentine
Stephen J
SJ
Predictive Physiology and Medicine, 1424 W. Adams Hill Circle, Bloomington, Indiana 47403, USA.
Plasencia
Manolo D
MD
Liu
Xiaoyun
X
Krishnan
Meera
M
Naylor
Stephen
S
Udseth
Harold R
HR
Smith
Richard D
RD
Clemmer
David E
DE
D001798:Blood Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002849:Chromatography, Gas / Q000295:instrumentation / Q000379:methods; D020869:Gene Expression Profiling; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D001798:Blood Proteins