There are over 100 scientific papers published on the advantages of the PCT platform, many by key opinion leaders worldwide. Such advantages include: (a) extraction and recovery of more membrane proteins, (b) enhanced protein digestion, (c) differential lysis in a mixed sample base, (d) pathogen inactivation, (e) increased DNA detection and (f) exquisite sample preparation process control.
Improved Proteolytic Digestion Under High Pressure Cycling: Rapid Digestion with Improved Sensitivity and Seque
Vera Gross1; John Wilson2; Alexander Lazarev1 1 Pressure BioSciences, Inc. Medford, MA 2 Protifi, LLC, Huntington
Comparative Lysis Efficiency of Escherichia coli
Comparative Lysis Efficiency of Escherichia coli at 20 and 35 kpsi Escherichia coli Lysis Rate with Varying Pressure and Cell Count Escherichia coli Lysis Rate with Multiple Passes and Pressures
Booth Poster: PCT-SWATH Work Flow with the Barocycler 2320EXT
~4 Hours from Tissue to Peptides PCT-SWATH was developed by PBI scientists and engineers in collaboration with Professor Ruedi Aebersold and Dr. Tiannan Guo of the Institute of Molecular Systems Biology, ETH Zurich, and the University of Zurich, Zurich, Switzerland
Compatibility of Deoxycholate Detergent with Pressure-enhanced Digestion by Trypsin and Lys-C: Alternative PCT-HD Workflow
High hydrostatic pressure accelerates protein digestion via a combination of two mechanisms. The first is the pressure-induced partial denaturation/unfolding and hydration of the target proteins, which leads to better access of the enzyme to its target sites. The second mechanism is less well understood, but relies on the increase in enzyme activity observed for some enzymes at elevated pressure, possibly resulting from the effect of pressure on hydrolysis. The combination of the two pressure-based mechanisms, one acting on the enzyme and the other acting on the substrates, leads to significantly accelerated digestion, but may need to be optimized for each enzyme and reagent system, to prevent pressure-induced denaturation and loss of activity of the enzyme itself. To this end, the effect of sodium deoxycholate (DOX) on PCT-accelerated trypsin and lys-C digestions, has been evaluated.
Carles Canto1 and Pablo M. Garcia-Roves2 1 Nestle Institute of Health Sciences S.A., Lausanne, Switzerland 2 Diabetes and Obesity Research Laboratory, Institut d'Investigacions Biomediques August Pi i Sunyer (IDIBAPS) and Spanish Biomedical Research Centre in Diabetes and Associated Metabolic Disorders (CIBERDEM), Barcelona, Spain
Introduction Proteomic profiling of mitochondria has the potential to provide insights into mitochondrial functions associated with aging, various metabolic states and diseases such as cancer, diabetes and cardiovascular disease . Rapid and reproducible isolation of intact mitochondria is crucial for efficient enrichment and subsequent proteomic analysis of low abundance mitochondrial proteins . Here we describe a system for the isolation of intact mitochondria from rat PC12 cells using pressure cycling technology (PCT).
Introduction Pressure cycling technology (PCT) has been proven to accelerate enzymatic protein digestion. For example, the effect of PCT on trypsin digestion has been demonstrated by several laboratories. They report that digestion times can be reduced from hours to minutes [1, 2]. Not only has PCT been shown to accelerate and improve protein digestion in solution, but it also can accelerate the digestion by trypsin of proteins in polyacrylamide gel slices . Additionally, the enhancing effect of PCT on the activity of several other enzymes, including Proteinase K, PNGase F, and Lys-C, has been reported [4, 5, 6]. It is thought that PCT may act synergistically with other protein denaturants, such as organic solvents and elevated temperature, to help maintain substrates in a denatured state leading to more exposure of enzyme target sites which results in better cleavage. Here we report the enhanced effect by PCT on the activity of the enzyme lysozyme. Lysozyme acts to hydrolyze peptidoglycans found in bacterial cell walls. This enzyme is frequently used for bacterial cell lysis prior to extracting DNA or proteins from bacteria. We also propose an alternate or additional mechanism by which PCT may enhance the activity of lysozyme.
Edmund Ting, Sc.D., Pressure BioSciences