title: Xenon hydrate dissociation measurements with model protein systems.
abstract: Effective long-term storage remains a significant challenge to the use and development of protein pharmaceuticals. We have investigated the interactions between clathrate hydrates and model protein solutions to determine the effects on hydrate formation. Here, the dissociation curve and equilibrium conditions for xenon clathrate hydrate with model lysozyme and lactate dehydrogenase (LDH) protein solutions have been studied using calorimetry measurements at pressures ranging from 3 to 20 bar. Sucrose in solution was shown to exhibit small inhibition effects on xenon hydrate formation, shifting the dissociation curve and decreasing the conversion of water to hydrate by 15-26%. The addition of l-histidine buffer and lysozyme at low concentrations did not substantially inhibit hydrate formation. However, small shifts in the dissociation curve were demonstrated for solutions containing LDH. The presence of lysozyme and LDH in solution did not significantly alter the conversion of water to hydrate, indicating that these and similar proteins do not substantially affect the extent of xenon gas hydrate formation. Preliminary experiments were performed for LDH solutions to assess the impact of xenon hydrate formation and dissociation on enzymatic activity, with samples stored in hydrate systems showing small decreases in activity.
authors.affiliation: Center for Hydrate Research, Chemical and Biological Engineering Department, Colorado School of Mines, Golden, Colorado 80401, USA.
authors.forename: Ryan D
authors.initials: RD
authors.lastname: Booker
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Koh
authors.forename: E Dendy
authors.initials: ED
authors.lastname: Sloan
authors.forename: Amadeu K
authors.initials: AK
authors.lastname: Sum
authors.forename: Evgenyi
authors.initials: E
authors.lastname: Shalaev
authors.forename: Satish K
authors.initials: SK
authors.lastname: Singh
mesh_terms: D000818:Animals; D004356:Drug Storage; D004795:Enzyme Stability; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry*; D011312:Pressure; D011817:Rabbits; D044366:Transition Temperature; D014867:Water / Q000737:chemistry*; D014978:Xenon / Q000737:chemistry*
chemical_list: D014867:Water; D014978:Xenon; D007770:L-Lactate Dehydrogenase; D009113:Muramidase