Abstract
Reducing the complexity of plasma proteome through complex multidimensional fractionation protocols is critical for the detection of low abundance proteins that have the potential to be the most specific disease biomarkers. Therefore, we examined a four dimension profiling method, which includes low abundance protein enrichment, tryptic digestion and peptide fractionation by IEF, SCX and RP-LC. The application of peptide pI filtering as an additional criterion for the validation of the identifications allows to minimize the false discovery rate and to optimize the best settings of the protein identification database search engine. This sequential approach allows for the identification of low abundance proteins, such as angiogenin (10−9 g/L), pigment epithelium growth factor (10−8 g/L), hepatocyte growth factor activator (10−7 g/L) and thrombospondin-1 (10−6 g/L), having concentrations similar to those of many other growth factors and cytokines involved in disease pathophysiology.
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Millioni, R., Tolin, S., Fadini, G.P. et al. High confidence and sensitivity four-dimensional fractionation for human plasma proteome analysis. Amino Acids 43, 2199–2202 (2012). https://doi.org/10.1007/s00726-012-1267-1
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DOI: https://doi.org/10.1007/s00726-012-1267-1