蛋白质-聚氨基酸沉淀可稳定治疗性蛋白质L-天冬酰胺酶，使其免受物理化学应激影响。
Protein-poly(amino acid) precipitation stabilizes a therapeutic protein l-asparaginase against physicochemical stress.
作者信息
Maruyama Takuya, Izaki Shunsuke, Kurinomaru Takaaki, Handa Kenji, Kimoto Tomoaki, Shiraki Kentaro
机构信息
Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan.
Research and Development Center, Terumo Corporation, 1500 Inokuchi, Nakai-machi, Ashigarakamigun, Kanagawa 259-0151, Japan.
出版信息
J Biosci Bioeng. 2015 Dec;120(6):720-4. doi: 10.1016/j.jbiosc.2015.04.010. Epub 2015 May 13.
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.
治疗性蛋白质的实际应用需要在水溶液中进行长期储存。最近，有人提出了一种沉淀-再溶解方法来制备盐可解离的蛋白质-聚电解质复合物（PPC）。为了阐明该复合物在蛋白质储存中的效用，我们研究了含有L-天冬酰胺酶（ASNase）和聚-L-赖氨酸（polyK）的PPC沉淀物的应激耐受性。通过沉淀-再溶解方法制备了含有ASNase和polyK的PPC沉淀物。将样品进行三种类型的应激处理，即加热、振荡和氧化。应激处理后测量样品上清液的蛋白质浓度、酶活性和圆二色光谱。与天然溶液相比，由ASNase和polyK组成的PPC沉淀物表现出对热应激和振荡应激的耐受性。此外，PPC沉淀物保护ASNase不被氧化失活。ASNase/polyK复合物的PPC沉淀物成功地使ASNase在物理化学应激下稳定。这些结果表明，PPC沉淀物作为不稳定蛋白质在水溶液中的储存方法具有很大的潜力。
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