%0 Journal Article
%T Spectral counting assessment of protein dynamic range in cerebrospinal fluid following depletion with plasma-designed immunoaffinity columns
%A Jacques Borg
%A Alex Campos
%A Claudio Diema
%A N¨²ria Ome£¿aca
%A Eliandre de Oliveira
%A Joan Guinovart
%A Marta Vilaseca
%J Clinical Proteomics
%D 2011
%I BioMed Central
%R 10.1186/1559-0275-8-6
%X We applied LC-MS/MS to compare the performance of two CSF enrichment techniques that immunodeplete either albumin alone (IgYHSA) or 14 high-abundance proteins (IgY14). In order to estimate dynamic range of proteins identified, we measured protein abundance with APEX spectral counting method.Both immunodepletion methods improved the number of low-abundance proteins detected (3-fold for IgYHSA, 4-fold for IgY14). The 10 most abundant proteins following immunodepletion accounted for 41% (IgY14) and 46% (IgYHSA) of CSF protein content, whereas they accounted for 64% in non-depleted samples, thus demonstrating significant enrichment of low-abundance proteins. Defined proteomics experiment metrics showed overall good reproducibility of the two immunodepletion methods and MS analysis. Moreover, offline peptide fractionation in IgYHSA sample allowed a 4-fold increase of proteins identified (520 vs. 131 without fractionation), without hindering reproducibility.The novelty of this study was to show the advantages and drawbacks of these methods side-to-side. Taking into account the improved detection and potential loss of non-target proteins following extensive immunodepletion, it is concluded that both depletion methods combined with spectral counting may be of interest before further fractionation, when searching for CSF biomarkers. According to the reliable identification and quantitation obtained with APEX algorithm, it may be considered as a cheap and quick alternative to study sample proteomic content.Biomarkers are key tools for detecting and monitoring neurodegenerative processes. Clinical Proteomics is especially well-suited to the discovery and implementation of biomarkers derived from biofluids. A major limiting factor for in-depth proteomics profiling is the immense dynamic range of biofluid proteins, which spans 10 to 12 orders of magnitude [1]. In human plasma, the 22 most abundant proteins are responsible for ~99% of the bulk mass of the total proteins, thus lea
%K CSF
%K APEX
%K Biomarkers
%K depletion column
%K enrichment
%K low-abundance proteins
%U http://www.clinicalproteomicsjournal.com/content/8/1/6