Open AccessAutomated 2D Peptide Separation on a 1D Nano-LC-MS System
Author(s) -
Paul Taylor,
Peter Nielsen,
Morten Beck Trelle,
Ole B. Hørning,
Michael B. Andersen,
Ole Vorm,
Michael F. Moran,
Thomas Kislinger
Publication year - 2009
Publication title -
journal of proteome research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.644
H-Index - 161
eISSN - 1535-3907
pISSN - 1535-3893
DOI - 10.1021/pr800986c
Subject(s) - proteome , chromatography , mass spectrometry , peptide , resolution (logic) , proteomics , chemistry , high resolution , instrumentation (computer programming) , chromatographic separation , high performance liquid chromatography , computer science , artificial intelligence , biochemistry , gene , geology , operating system , remote sensing
Given the complexity of the mammalian proteome, high-resolution separation technologies are required to achieve comprehensive proteome coverage and to enhance the detection of low-abundance proteins. Among several technologies, Multidimensional Protein Identification Technology (MudPIT) enables the on-line separation of highly complex peptide mixtures directly coupled with mass spectrometry-based identification. Here, we present a variation of the traditional MudPIT protocol, combining highly sensitive chromatography using a nanoflow liquid chromatography system (nano-LC) with a two-dimensional precolumn in a vented column setup. When compared to the traditional MudPIT approach, this nanoflow variation demonstrated better first-phase separation leading to more proteins being characterized while using rather simple instrumentation and a protocol that requires less time and very little technical expertise to perform.
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