Rapid Identification and Characterization of Tryptic Peptides Using High Linear Velocity Nanobore UPLC MALDI MS/MS and Ion Mobility Separation

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Jean-Marie Casanova, Christian Claude, Marten Snel, Emmanuelle Claude, Mark McDowall, Iain Campuzano, Therese McKenna, James Langridge
Spectrometric de Masse et Analyse Proteomique, September 17 - 20, 2007, Pau France
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Here we describe the use of elevated flow rates combined with nanoscale columns packed with 1.7µm particles for rapid peptide based separations using a high pressure nanoUPLC system. Increasing the flow rate of the separation from approximately 0.3µL/min to 1.4µL/min has the effect of increasing the pressure of the chromatographic system from 1950psi  to 5900psi which allows the Chromatographic run time to be compressed to 8 minutes injection to injection.  Conventional HPLC run times for this type of experiment are typically 45 minutes to 1 hour. The eluent is combined with matrix solution and spotted directly onto MALDI target plates using a spotting robot. This combined with an orthogonal acceleration time-of-flight mass spectrometer equipped with a 200Hz repetition rate Nd:YAG laser allows for the rapid characterisation of simplified protein mixtures.  A novel ion mobility separation device was used to separate isobaric peptides.

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