Studying the proteome of blood serum was a natural fit for scientists at PNNL, which has a strong proteomics capability. A proteome is the collection of proteins expressed by a cell under a specific set of conditions at a certain time. Through its Biomolecular Systems Initiative, the laboratory is supporting multidisciplinary research in systems biology. Scientists have developed unique technologies that allow for more thorough analysis of proteins and have studied the proteome of ovarian cancer as well as other disease states.
Pounds and his team, which included lead author and post-doctoral researcher Joshua Adkins, used chromatography and mass spectrometry instead of the more traditional 2-D gel electrophoresis to identify proteins, including low-abundance proteins not previously identified in serum and proteins with an unknown function. Their overall analysis was conducted on a single human blood serum sample from a healthy anonymous female donor.
The majority of serum protein consists of a few, very abundant proteins. One of the current challenges in the field is that the presence of abundant proteins obscures the measurement of many low-abundance proteins, and that removal of these abundant proteins may result in the simultaneous removal of low-abundance proteins. Here, Pounds and his team kept those abundant proteins, but simplified the mass spectrometry by fractionating the peptides according to charge state.
Once fractionated to allow for the analysis of lower abundance proteins, the samples were analyzed using a mass spectrometer that had been programmed to concentrate on specific ranges of peptide size during several analyses, thereby providing a more complete analysis of the proteome. The researchers employed powerful mas
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Contact: Staci Maloof
staci.maloof@pnl.gov
509-372-6313
DOE/Pacific Northwest National Laboratory
18-Dec-2002