Application of pressurized solvents for ultrafast trypsin hydrolysis in proteomics: Proteomics on the fly

Journal Article · Fri Aug 01 00:00:00 EDT 2008 · Journal of Proteome Research, 7(8):3276-3281

DOI:https://doi.org/10.1021/pr7008077· OSTI ID:949109

Lopez-Ferrer, Daniel; Petritis, Konstantinos; Hixson, Kim K.; Heibeck, Tyler H.; Moore, Ronald J.; Belov, Mikhail E.; Camp, David G.; Smith, Richard D.

A new method for rapid proteolytic digestion of proteins under high pressure that uses pressure cycling technology in the range of 5 to 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.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 949109
• Report Number(s): PNNL-SA-60252; 400412000
• Journal Information: Journal of Proteome Research, 7(8):3276-3281, Journal Name: Journal of Proteome Research, 7(8):3276-3281 Journal Issue: 8 Vol. 7
• Country of Publication: United States
• Language: English