Proteomic analysis of human blood serum using peptide library beads.
Human serum is thought to contain key information for diagnostics of human disease. However, no single technology is currently nor might ever be available to cope with the complexity and dynamic range of the serum proteome. We here report a large-scale proteomic study of human blood serum using peptide library beads and mass spectrometry. Serum proteins are adsorbed onto polymeric beads coated with a combinatorial library composed of millions of hexameric peptide baits. Analysis of the eluates from this combinatorial library (as obtained with 3 eluants of different strength, able to release 99% of the retentate) via liquid chromatography coupled to high-resolution mass spectrometry resulted in the identification of 1559 proteins or 3869 proteins, respectively, depending on how 95% confidence was estimated. In either case, the analysis showed that ligand beads are able to capture a large number of proteins in a single operation. The ligand bead bound fraction appeared to have a lower dynamic range when compared to the starting material, due to a "normalization" of the protein concentrations in the original mixture. We find that extensive information on the protein composition of complex samples such as serum can be obtained using ligand beads and that these beads enrich the proteomic tool box.
Sennels
Lau
L
The FIRC Institute for Molecular Oncology Foundation, Via Adamello 16, I-20139 Milan, Italy.
Salek
Mogjiborahman
M
Lomas
Lee
L
Boschetti
Egisto
E
Righetti
Pier Giorgio
PG
Rappsilber
Juri
J
D001798:Blood Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008024:Ligands; D013058:Mass Spectrometry / Q000379:methods; D008863:Microspheres; D009842:Oligopeptides / Q000737:chemistry; D019151:Peptide Library; D011109:Polymethacrylic Acids / Q000737:chemistry; D020543:Proteome / Q000032:analysis*; D044967:Serum
D001798:Blood Proteins; D008024:Ligands; D009842:Oligopeptides; D019151:Peptide Library; D011109:Polymethacrylic Acids; D020543:Proteome