Rosa Viner1; Leigh A Foster2; Ryan D. Bomgarden2; Michael W. Belford1; Satendra Prasad1; Romain Huguet1; Eloy R. Wouters1
ASMS 2019 Poster
Purpose: To evaluate widely used enrichment/fractionation techniques and the newly developed High Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS)1 device for cross-linked peptide analysis using a Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ mass spectrometer.
Methods: Different amine-reactive, homobifuctional crosslinkers were used to crosslink bovine serum albumin (BSA), yeast enolase, human cell lysate and mouse mitochondria. Crosslinked samples were reduced, alkylated and digested with trypsin before MS analysis. Cross-linked peptides were pre-fractionated on SCX or peptide size exclusion chromatography (SEC) columns. An Orbitrap Fusion Lumos mass spectrometer with/out the Thermo Scientific™ FAIMS Pro™ Interface was used for crosslinked peptide analysis. Data analysis was performed with Thermo Scientific™ Proteome Discoverer™ 2.3 software using a XlinkX 2.0 software node.
Results: FAIMS alone or in combination with enrichment/pre-fractionation significantly improves identification of cross-linked peptides in complex samples.
1Thermo Fisher Scientific, San Jose, CA; 2Thermo Fisher Scientific, Rockford, IL
United States
