Toward a Human Blood Serum Proteome

By J. Adkins, S. Varnum, K. Auberry, Ronald J. Moore, N. Angell, Richard D. Smith, D. Springer, J. Pounds. Molecular & Cellular Proteomics, 2002

758 citations recorded by the source corpus.

Blood serum is a complex body fluid that contains various proteins ranging in concentration over at least 9 orders of magnitude. Using a combination of mass spectrometry technologies with improvements in sample preparation, we have performed a proteomic analysis with submilliliter quantities of serum and increased the measurable concentration range for proteins in blood serum beyond previous reports. We have detected 490 proteins in serum by on-line reversed-phase microcapillary liquid chromatography coupled with ion trap mass spectrometry. To perform this analysis, immunoglobulins were removed from serum using protein A/G, and the remaining proteins were digested with trypsin. Resulting peptides were separated by strong cation exchange chromatography into distinct fractions prior to analysis. This separation resulted in a 3–5-fold increase in the number of proteins detected in an individual serum sample. With this increase in the number of proteins identified we have detected some lower abundance serum proteins (ng/ml range) including human growth hormone, interleukin-12, and prostate-specific antigen. We also used SEQUEST to compare different protein databases with and without filtering. This comparison is plotted to allow for a quick visual assessment of different databases as a subjective measure of analytical quality. With this study, we have performed the most extensive analysis of serum proteins to date and laid the foundation for future refinements in the identification of novel protein biomarkers of disease.

Subfields: Molecular Biology, Spectroscopy

View Paper: https://www.semanticscholar.org/paper/d08910093a7777bd896c404ee661257ce7bdd296

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Citations in the corpus

Proteomics: Technologies and Their Applications. (2017, background): “PSA, human growth hormone and interleukin-12 were also analyzed from human serum (124).”
The Human Plasma Proteome (2004, methodology): “The three experimental datasets are derived from 1) whole protein separation by a three-dimensional process (immunosubtraction/ion exchange/size exclusion) followed by two-dimensional electrophoresis (2DE) followed by MS identification of resolved spots (4); 2) Ig subtraction followed by trypsin dig”
Quantitative Mass Spectrometric Multiple Reaction Monitoring Assays for Major Plasma Proteins* (2006, background): “(23) by counting the number of separate “hits” for the peptide in the data set divided by the number of hits for the most frequently detected peptide from the same protein.”
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Identification of Proteins in Human Cytomegalovirus (HCMV) Particles: the HCMV Proteome (2004, methodology): “Although gM complexes with UL73 (gN), only 0.1% of the peptide content detected in our virion preparations was gN. Lastly, UL75 (gH) and UL115 (gL), which a Asterisks indicate proteins whose LC-MS/MS SEQUEST scores were lower than our cutoff values; however, the proteins were identified with the mor”
Enzyme Biosensors for Biomedical Applications: Strategies for Safeguarding Analytical Performances in Biological Fluids (2016, neutral): “[135].”
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Characterization of the Low Molecular Weight Human Serum Proteome*S (2003, methodology): “The inclusion of these viral sequences in the database has the same effect as searching the data against databases from other organisms (9) or other approaches such as searching against reversed databases (16).”
Plasma Proteome Database as a resource for proteomics research: 2014 update (2013, methodology): “used two different separation techniques followed by mass spectrometry (MS) for characterization of proteins from depleted serum and reported 490 proteins (20).”
The Human Plasma Proteome (2002, methodology): “Recent values of 250 (*) and 490 ( ) proteins come from LC/LC/2-DE (see Footnote 3) and LC/LC/MS/MS (70), respectively.”