|
Home > Magazine > Liquid Chromatography
Faster LC Sample PrepPressure cycling technology introduces a new thermodynamic dimension in sample preparation for HPLC and LC-MS analyses. by Alexander Lazarev, PhD, Pressure BioSciences, Inc.
Since Blaise Pascal in the mid-seventeenth century described physical laws governing the effects of pressure on matter, the laws of physics haven’t changed. However, until recently, the adoption of high hydrostatic pressure equipment in the lab has been slow due to the lack of materials and engineering solutions suitable to generate and maintain high pressure in a benchtop instrument.
To meet this need, Pressure Biosciences developed ultra-high pressure instruments based on proprietary pressure cycling technology (PCT). PCT destabilizes inter-molecular interactions by rapidly and repeatedly raising and lowering pressure in the reaction vessel from ambient to 35,000 psi.
PULSE (left) and PCT microtubes accept 50- to 150-mL samples. | High hydrostatic pressure acts preferentially on the more compressible constituents of the sample, such as lipids and proteins. Thus, selective energy distribution results in destabilization of molecular interactions in the lipid bilayers and other cellular components, but not in the disruption of covalent bonds. PCT instruments (Barocycler NEP-3229/2320) are described as unique thermodynamic lab tools that provide control of both temperature and pressure for various chemical processes.
Applications of PCT instrumentation include cell and tissue lysis, dissolution and fractionation of biological macromolecules by their solubility and cellular localization, exquisite control of molecular interactions, and accelerated enzymatic digestion of protein samples for proteomics.
Tissue disruption with The PCT Shredder combines rapid and convenient mechanical tissue disruption, resulting in a convenient, single-tube method for preparing whole tissue lysates from tough samples ranging from plant tissue to cardiac and skeletal muscle.
Pressure BioSciences also developed a detergent-free sample prep technique that allows isolation of protein from lipid-rich samples such as adipose tissue, brain tissue and membrane preparations. This method takes advantage of a synergistic combination of sample disruption by alternating hydrostatic pressure, pressure cycling technology, and a reagent system provided in the ProteoSolve-SB kit that dissolves and partitions proteins, lipids and nucleic acids into separate fractions from a single tissue sample, enabling analysis of single small-tissue samples.
PBI is developing a flow-through model of the Barocycler, which would allow direct integration of PCT with HPLC. Such integration could enable reproducible, unattended online enzymatic digestion of protein samples in LC-MS-MS proteomic experiments.
For more information, call 508-230-1828 or visit www.pressurebiosciences.com
|
|
|
|
|
|
|