| Proteomics under pressure. Development of essential sample preparation techniques in proteomics using ultra-high hydrostatic pressure. | |
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MedLine Citation:
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PMID: 22029901 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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The minimization of pre-analytical variables in sample preparation is imperative for successful discovery-driven and translational research involving large-scale biomolecular profiling. Here we demonstrate a novel technique using high hydrostatic pressure in addition to several chaotropes and solvents to maximize efficiency of both cell lysis and enzymatic digestion while minimizing the time, manual involvement in sample processing, and pre-analytical variability introduced prior to mass spectrometry-based proteomics analysis. The digestion techniques were evaluated and optimized for in-solution, in-gel, and on-membrane applications using protein standards and cell lysates. The lysis techniques were evaluated using human HepG2 cells. Our results demonstrate that the use of elevated pressure and organic solvents can achieve superior protein recovery of organelle-, complex-, and especially membrane-associated proteins, meanwhile obtaining more than a 20-fold increase in throughput with improved reproducibility. This study introduces the concept of <i>ultra-high performance sample preparation platforms</i> for targeted characterization of proteome subsets in biological systems. |
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Authors:
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Emily Freeman; Alexander R Ivanov |
Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-10-26 |
Journal Detail:
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Title: Journal of proteome research Volume: - ISSN: 1535-3907 ISO Abbreviation: - Publication Date: 2011 Oct |
Date Detail:
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Created Date: 2011-10-27 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101128775 Medline TA: J Proteome Res Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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