Journal
EXPERT REVIEW OF PROTEOMICS
Volume 8, Issue 6, Pages 787-800Publisher
EXPERT REVIEWS
DOI: 10.1586/EPR.11.67
Keywords
bottom-up MS; gel electrophoresis; gel-free separation; protein solubilization; proteome fractionation; SDS removal; surfactants; top-down MS
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Funding
- National Sciences and Engineering Research Council of Canada
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Proteome fractionation refers to separation at the level of intact proteins. Proteome fractionation may precede sample digestion and subsequent peptide-level separation and detection (i.e., bottom-up mass spectrometry [MS]). For top-down MS, proteome fractionation acts as a stand-alone separation platform, since intact proteins are directly analyzed by the mass spectrometer. Regardless of the MS identification strategy, separation of intact proteins has clear benefits as a result of decreasing sample complexity. However, this stage of the workflow also creates considerable challenges, which are generally absent from the counterpart peptide separation experiment. For example, maintaining protein solubility is a key concern before, during and after separation. To this end, surfactants such as sodium dodecyl sulfate may be employed during fractionation, so long as they are eliminated prior to MS. In this article, current strategies for proteome fractionation in a MS-compatible format are reviewed, illustrating the challenges and outlooks on this important aspect of proteomics.
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