Towards defining the urinary proteome using liquid chromatography-tandem mass spectrometry. II. Limitations of complex mixture analyses.

Abstract:

:With an emphasis on obtaining a multitude of high quality tandem mass spectrometry spectra for protein identification, instrumental parameters are described for the liquid chromatography-tandem mass spectrometry analysis of trypsin digested unfractionated urine using a hybrid quadrupole-time-of-flight (Q-TOF) mass spectrometer. Precursor acquisition rates of up to 20 distinct precursors/minute in a single analysis were obtained through the use of parallel precursor selection (four precursors/survey period) and variable collision induced dissociation integration time (1 to 6 periods summed). Maximal exploitation of the gas phase fractionated ions was obtained through the use of narrow survey scans and iterative data-dependent analyses incorporating dynamic exclusion. The impact on data fidelity as a product of data-dependent selection of precursor ions from a dynamically excluded field is discussed with regards to sample complexity, precursor selection rates, survey scan range and facile chemical modifications. Operational and post-analysis strategies are presented to restore data confidence and reconcile the greatest number of matched spectra.

journal_name

Proteomics

journal_title

Proteomics

authors

Davis MT,Spahr CS,McGinley MD,Robinson JH,Bures EJ,Beierle J,Mort J,Yu W,Luethy R,Patterson SD

doi

10.1002/1615-9861(200101)1:1<108::AID-PROT108>3.0.

subject

Has Abstract

pub_date

2001-01-01 00:00:00

pages

108-17

issue

1

eissn

1615-9853

issn

1615-9861

journal_volume

1

pub_type

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