Abstract
Over the years many methodologies have been advanced to aid in the enhancement of proteolysis cleavage efficiency, reaction time, and protein sequence coverage for LC‐MS/MS based analysis. The series of experiments presented here examine the effect pressure cycling on each of these factors. Several individual proteins and a simple protein mixture were digested with different proteases under high pressure using a Barocycler NEP2320 pressure cycler and compared to overnight at 1 atm and 37 °C proteolysis conditions. The resulting peptides were identified and quantified using an LC‐MS/MS workflow with a LTQ XL with ETD. The results indicate that digestions using pressure cycling produce more distinct peptides and equivalent or higher protein sequence coverage when compared to standard overnight digestions.
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