Use of peptide analogue diversity library beads for increased depth of proteomic analysis: application to cerebrospinal fluid.
Biological samples can contain proteins with concentrations that span more than 10 orders of magnitude. Given the limited dynamic range of analysis methods, observation of proteins present at the lower concentrations requires depletion of high-abundance proteins, or other means of reducing the dynamic range of concentrations. Hexapeptide diversity library beads have been used to bind proteins in a complex sample up to a given saturation limit, effectively truncating the maximum concentration of proteins at a desired level. To avoid the potential problem of susceptibility of the hexapeptides to cleavage by proteases in the sample and/or bacterial degradation, peptide analogues that exhibit similar binding characteristics to peptides can be used in place of peptides. We report here the use of hexameric peptoid diversity library beads to reduce the dynamic range of protein concentrations in human cerebrospinal fluid (CSF). Using this method in conjunction with 2D LC/MS/MS analyses, we identified 200 unique proteins, about twice the number identified in untreated CSF.
Shores
Kevin S
KS
Department of Pharmacology and MUSC Proteomics Center, Medical University of South Carolina, Charleston, SC 29425, USA.
Udugamasooriya
D Gomika
DG
Kodadek
Thomas
T
Knapp
Daniel R
DR
D002555:Cerebrospinal Fluid / Q000737:chemistry*; D020650:Combinatorial Chemistry Techniques; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019151:Peptide Library; D010455:Peptides / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D020543:Proteome / Q000032:analysis*
D019151:Peptide Library; D010455:Peptides; D020543:Proteome