Proteomics of Cerebrospinal Fluid: Throughput and Robustness Using a Scalable Automated Analysis Pipeline for Biomarker Discovery.

摘要: Cerebrospinal fluid (CSF) is a body of high clinical relevance and an important source potential biomarkers for brain-associated damages, such as traumatic brain injury stroke, diseases, Alzheimer’s Parkinson’s. Herein, we have implemented, evaluated, validated scalable automated proteomic pipeline (ASAP2) the sample preparation analysis CSF, enabling increased throughput robustness biomarker discovery. Human CSF samples were depleted from abundant proteins subjected to reduction, alkylation, protein digestion, tandem mass tag (TMT) 6-plex labeling, pooling, cleanup in 96-well-plate format before reversed-phase liquid chromatography spectrometry (RP-LC MS/MS). We showed impact on proteome coverage applying depletion proteins, which usually performed blood plasma or serum samples. Using ASAP2 analyze 96 identical samples, determined analytical f...