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Application of Pressurized Solvents for Ultrafast Trypsin Hydrolysis in Proteomics: Proteomics on the Fly

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journal contribution
posted on 01.08.2008, 00:00 by Daniel López-Ferrer, Konstantinos Petritis, Kim K. Hixson, Tyler H. Heibeck, Ronald J. Moore, Mikhail E. Belov, David G. Camp, Richard D. Smith
A new method for rapid proteolytic digestion of proteins under high pressure that uses pressure cycling technology in the range of 5−35 kpsi was demonstrated for proteomic analysis. Successful in-solution digestions of single proteins and complex protein mixtures were achieved in 60 s and then analyzed by reversed phase liquid chromatography-electrospray ionization ion trap-mass spectrometry. Method performance in terms of the number of Shewanella oneidensis peptides and proteins identified in a shotgun approach was evaluated relative to a traditional “overnight” sample preparation method. Advantages of the new method include greatly simplified sample processing, easy implementation, no cross contamination among samples, and cost effectiveness.