title: Freeze-drying of proteins with glass-forming oligosaccharide-derived sugar alcohols.
abstract: Physical properties and protein-stabilizing effects of sugar alcohols in frozen aqueous solutions and freeze-dried solids were studied. Various frozen sugar alcohol solutions showed a glass transition of the maximally freeze-concentrated phase at temperatures (T(g)'s) that depended largely on the solute molecular weights. Some oligosaccharide-derived sugar alcohols (e.g., maltitol, lactitol, maltotriitol) formed glass-state amorphous cake-structure freeze-dried solids. Microscopic observation of frozen maltitol and lactitol solutions under vacuum (FDM) indicated onset of physical collapse at temperatures (T(c)) several degrees higher than their T(g)'s. Freeze-drying of pentitols (e.g., xylitol) and hexitols (e.g., sorbitol, mannitol) resulted in collapsed or crystallized solids. The glass-forming sugar alcohols prevented activity loss of a model protein (LDH: lactate dehydrogenase) during freeze-drying and subsequent storage at 50 degrees C. They also protected bovine serum albumin (BSA) from lyophilization-induced secondary structure perturbation. The glass-forming sugar alcohols showed lower susceptibility to Maillard reaction with co-lyophilized L-lysine compared to reducing and non-reducing disaccharides during storage at elevated temperature. Application of the oligosaccharide-derived sugar alcohols as alternative stabilizers in lyophilized protein formulations was discussed.
authors.affiliation: Faculty of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Kadoya
authors.forename: Kahori
authors.initials: K
authors.lastname: Fujii
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Izutsu
authors.forename: Etsuo
authors.initials: E
authors.lastname: Yonemochi
authors.forename: Katsuhide
authors.initials: K
authors.lastname: Terada
authors.forename: Chikako
authors.initials: C
authors.lastname: Yomota
authors.forename: Toru
authors.initials: T
authors.lastname: Kawanishi
mesh_terms: D000818:Animals; D002417:Cattle; D002626:Chemistry, Pharmaceutical / Q000379:methods; D004355:Drug Stability; D004356:Drug Storage; D005079:Excipients / Q000737:chemistry*; D005612:Freeze Drying; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D008970:Molecular Weight; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013402:Sugar Alcohols / Q000737:chemistry*; D013696:Temperature; D044366:Transition Temperature
chemical_list: D005079:Excipients; D013402:Sugar Alcohols; D012710:Serum Albumin, Bovine; D007770:L-Lactate Dehydrogenase