title: Protein instability toward organic solvent/water emulsification: implications for protein microencapsulation into microspheres.
abstract: The objective of this study was to investigate the behavior of three proteins at an organic solvent/water interface. To simulate the first microencapsulation step of a water-in-oil-in-water emulsion technique, a water-in-oil emulsion was prepared by emulsifying an aqueous protein solution in either methylene chloride or ethyl acetate. Phase separation was then followed to collect protein samples from the aqueous phase and the organic solvent/water interface. Their properties were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size exclusion-HPLC. Bovine serum albumin was relatively unharmed during emulsification, compared to other proteins such as ovalbumin and lysozyme. In particular, the methylene chloride treatment on ovalbumin led to the formation of a large quantity of water-insoluble, solid-like aggregates and changes in the composition of monomeric and dimeric ovalbumin species. With regard to the question of ovalbumin recovery, only 9.74 approximately 37.72% of the used ovalbumin was present in the aqueous phases after emulsification. Similar penchant was noted with lysozyme. Water-insoluble aggregates brought with by emulsification were found to be covalently bound. Interestingly, less emulsification-induced denaturing effects were observed with ethyl acetate. Our study clearly demonstrated the emulsification-induced adverse events that were detrimental to the integrity of proteins and the importance of preserving protein stability toward microencapsulation.
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee College of Pharmacy, Memphis, USA.
authors.forename: H
authors.initials: H
authors.lastname: Sah
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004339:Drug Compounding; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004655:Emulsions; D008863:Microspheres; D011506:Proteins / Q000737:chemistry*; D012997:Solvents; D014867:Water
chemical_list: D004655:Emulsions; D011506:Proteins; D012997:Solvents; D014867:Water