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Anal Chem. 2011 Mar 15;83(6):2250-7. doi: 10.1021/ac103079q. Epub 2011 Feb 22.

Faster, quantitative, and accurate precursor acquisition independent from ion count.

Author information

1
Department of Medicinal Chemistry, University of Washington, Seattle, Washington, United States.

Abstract

Data-dependent precursor ion selection is widely used in shotgun proteomics to profile the protein components of complex samples. Although very popular, this bottom-up method presents major drawbacks in terms of detectable dynamic range. Recently, we demonstrated the superior performance of a data-independent method we termed precursor acquisition independent from ion count (PAcIFIC). Here, we report a faster, accurate, multiplexed, and quantitative PAcIFIC method. Our results show that the time needed to perform such analysis can be decreased by 33% to 66% using modern ion trap instruments and that high mass accuracy can be applied to such a strategy. Quantification capability is demonstrated on protein standards and a whole bacterial cell lysate using isobaric tagging. Finally, we confirm in yeast the dynamic range capabilities of such a method where proteins down to less than 50 copies per cell can be monitored without sample prefractionation.

PMID:
21341720
PMCID:
PMC3217585
DOI:
10.1021/ac103079q
[Indexed for MEDLINE]
Free PMC Article

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