期刊
JOURNAL OF PROTEOME RESEARCH
卷 7, 期 8, 页码 3276-3281出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr7008077
关键词
mass spectrometry; trypsin digestion; pressure cycling technology; bottom-up proteomics
资金
- NCI NIH HHS [R21 CA 12619-01, R33 CA126191, R33 CA126191-02] Funding Source: Medline
- NCRR NIH HHS [RR 018522, P41 RR018522, P41 RR018522-06] Funding Source: Medline
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.
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