title: Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment.
abstract: Reversible phosphorylation plays a key role in numerous biological processes. Mass spectrometry-based approaches are commonly used to analyze protein phosphorylation, but such analysis is challenging, largely due to the low phosphorylation stoichiometry. Hence, a number of phosphopeptide enrichment strategies have been developed, including metal oxide affinity chromatography (MOAC). Here, we describe a new material for performing MOAC that employs a magnetite-doped polydimethylsiloxane (PDMS), that is suitable for the creation of microwell array and microfluidic systems to enable low volume, high throughput analysis. Incubation time and sample loading were explored and optimized and demonstrate that the embedded magnetite is able to enrich phosphopeptides. This substrate-based approach is rapid, straightforward and suitable for simultaneously performing multiple, low volume enrichments.
authors.affiliation: Institute of Molecular, Cell and Systems Biology, University of Glasgow, Glasgow, G12 8QQ, UK.
authors.forename: Mairi E
authors.initials: ME
authors.lastname: Sandison
authors.forename: K Tveen
authors.initials: KT
authors.lastname: Jensen
authors.forename: F
authors.initials: F
authors.lastname: Gesellchen
authors.forename: J M
authors.initials: JM
authors.lastname: Cooper
authors.forename: A R
authors.initials: AR
authors.lastname: Pitt
mesh_terms: D000818:Animals; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques; D010748:Phosphopeptides / Q000032:analysis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002364:Caseins; D004129:Dimethylpolysiloxanes; D010748:Phosphopeptides; C013830:baysilon; D052203:Ferrosoferric Oxide