Dynamic range compression via hexapeptide libraries for increased proteome coverage in whole human saliva

摘要

Testing effect of protein dynamic range compression and peptide fractionation strategies on proteomic coverage of complex biological samples using shotgun proteomics. As an example of a complex biological sample, we used whole saliva which represents a biological fluid with considerable disease diagnostic potential (Figure 1). Fresh saliva was obtained from healthy volunteers, centrifuged at 16,000 g for 10 minutes, and clarified supernatant supplemented with protease inhibitors. Untreated saliva was boiled 10 minutes in equal volume of 2X SDS sample buffer, acetone precipitated, and dissolved in 50mM Tris, pH 8.0, 5mM EDTA, 0.5percent SDS, diluted ten-fold in same buffer lacking SDS, and trypsinized.