• Digesting proteins into peptides is an
essential step in bottom-up proteomics.
Recently, pressure assisted digestion using
pressure cycling technology (PCT) was
demonstrated to significantly speed time
needed for digestions from hours to
minutes.
• In this work, we developed a novel ultra-high
pressure PCT reactor that couples to an LCMS
platform. The reactor consists of a
pressure generator, a temperature
controlled flow-through pressure cell,
isolated from the flow path by high pressure
valves.
• The system was tested initially with a pool of
standard proteins and later with a set of
digestion resistant proteins, including the
hydrophobic membrane-spanning protein
bacteriorhodopsin (Halobacterium halobium)
and a water-insoluble fraction of Shewanella
oneidensis proteins.