Proteomic Analysis of Human Ventricular Cerebrospinal Fluid from Neurologically Normal, Elderly Subjects Using Two-Dimensional LC−MS/MS

• DOI: 10.1021/pr034070r
• Publication Date: 2004-02-09T05:19:25Z
• Authors: Brett R. Wenner; Mark A. Lovell; Bert C. Lynn
• Keywords: Proteome

A two-dimensional liquid chromatography separation scheme coupled to tandem mass spectrometry (2-D LC−MS/MS) was utilized profile the proteome of human CSF. Ventricular CSF samples acquired post-mortem from 10 cognitively normal elderly subjects (mean ± SEM Braak stage = 1.7 0.2) were analyzed determine their protein composition. Raw subjected an immuno-based processing method remove highly abundant albumin and immunoglobulin (Ig), allowing better detection lower-abundance proteins. Samples trypsin proteolysis followed by C18 solid-phase extraction. Tryptic peptides separated using a 2-D LC column, in which both strong cation exchange (SCX) phases packed into single capillary. MS/MS spectra searched against sub-database NBCI nonredundant database SEQUEST algorithm. Search results further filtered DTAselect, individual compared one another Contrast. Using this method, we able unambiguously identify 249 proteins subjects. Of these proteins, 38% unique subjects, whereas only 6% common all These suggest considerable subject-to-subject variability proteome. Keywords: cerebrospinal fluid • proteomics LC−MS/MS Alzheimer's Disease biomarkers