title: Protein-poly(amino acid) precipitation stabilizes a therapeutic protein l-asparaginase against physicochemical stress.
abstract: Long-term storage in aqueous solution has been demanded for the practical application of therapeutic proteins. Recently, a precipitation-redissolution method was proposed to prepare salt-dissociable protein-polyelectrolyte complex (PPC). To elucidate the utility of the complex for storage of proteins, we investigated the stress tolerance of PPC precipitates containing l-asparaginase (ASNase) and poly-l-lysine (polyK). PPC precipitate containing ASNase and polyK was prepared by precipitation-redissolution method. The sample was treated to three types of stress, i.e., heat, shaking, and oxidation. The protein concentration, enzyme activity, and CD spectrum of the supernatants of samples were measured after stressed. PPC precipitate consisting of ASNase and polyK showed tolerance against thermal and shaking stress compared to the native solution. In addition, PPC precipitate protected ASNase from inactivation by oxidation. PPC precipitate of ASNase/polyK complex successfully stabilized ASNase against physicochemical stresses. These results suggest that the PPC precipitate has great potential as a storage method in aqueous solution for unstable proteins.
authors.affiliation: Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Maruyama
authors.affiliation: Research and Development Center, Terumo Corporation, 1500 Inokuchi, Nakai-machi, Ashigarakamigun, Kanagawa 259-0151, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Izaki
authors.affiliation: Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kurinomaru
authors.affiliation: Research and Development Center, Terumo Corporation, 1500 Inokuchi, Nakai-machi, Ashigarakamigun, Kanagawa 259-0151, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Handa
authors.affiliation: Research and Development Center, Terumo Corporation, 1500 Inokuchi, Nakai-machi, Ashigarakamigun, Kanagawa 259-0151, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Kimoto
authors.affiliation: Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan. Electronic address: shiraki@bk.tsukuba.ac.jp.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Shiraki
mesh_terms: D001215:Asparaginase / Q000737:chemistry* / Q000378:metabolism; D011232:Chemical Precipitation; D004356:Drug Storage / Q000379:methods*; D004573:Electrolytes / Q000737:chemistry; D004789:Enzyme Activation; D006358:Hot Temperature; D009038:Motion; D010084:Oxidation-Reduction; D011107:Polylysine / Q000737:chemistry*; D055550:Protein Stability; D012995:Solubility; D012996:Solutions
chemical_list: D004573:Electrolytes; D012996:Solutions; D011107:Polylysine; D001215:Asparaginase
keywords: Precipitation; Protein-polyelectrolyte complex; Redissolution; Stabilization; l-Asparaginase