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Peptide profiling of biological samples using MALDI-TOF mass spectrometry (MS) is an increasingly popular approach used for the discovery of biomarkers and as a means to detect and diagnose disease and allow the assessment of disease severity, progression, and the effectiveness of treatments.
Before mass spectrometry can be used to generate body fluid peptide profiles, reproducible, preferably automated, sample preparation procedures need to be used in which peptides are enriched and substances which interfere with MS analysis removed. Dynabeads are uniform superpara-magnetic monodisperse beads with a specific and defined surface for the adsorption/desorption and coupling of bio-reactive molecules. We have developed ion exchange and reversed-phase magnetic beads for peptide isolation from body fluids. These beads enable large numbers of peptides to be processed and analyzed simultaneously. The use of multiple bead surfaces for the capture of peptides from the same sample increases the number of peptides detected in a given sample and may increase the likelihood of discovering novel biomarkers. Sample preparation procedures have been successfully automated on robotic platforms.
Using peptide capture automated on the KingFisher96 coupled to MALDI-TOF-MS read-out, we investigated pre-analytical variables and have identified peptides sensitive to differences in clotting time. Furthermore, we have optimized the protocol for capture of native CSF peptides.