Inulin glasses for the stabilization of therapeutic proteins

海藻糖 菊粉 玻璃化转变 化学 蔗糖 结晶 冷冻干燥 稳定器(航空) 碳水化合物 食品科学 色谱法 生物化学 有机化学 聚合物 机械工程 工程类
作者
Wouter L. J. Hinrichs,Maarten Prinsen,Henderik W. Frijlink
出处
期刊:International Journal of Pharmaceutics [Elsevier BV]
卷期号:215 (1-2): 163-174 被引量:157
标识
DOI:10.1016/s0378-5173(00)00677-3
摘要

Sugar glasses are widely used to stabilize proteins during drying and subsequent storage. To act successfully as a protectant, the sugars should have a high glass transition temperature (Tg), a poor hygroscopicity, a low crystallization rate, and contain no reducing groups. When freeze drying is envisaged as method of drying, a relatively high Tg of the freeze concentrated fraction (Tg') is preferrable. In this study, whether inulins meet these requirements was investigated. Inulins of various degrees of polymerisation (DP) were evaluated. Trehalose glass was used as a positive control. It was found that the Tg and the Tg' of inulins with a number/weight average DP (DP(n)/DP(w)) higher than 5.5/6.0 were higher than those of trehalose glass. Furthermore, inulin glasses showed a similar hygroscopicity to that of trehalose glass but crystallized less rapidly. Less than 6% of the sugar units of inulins with a DP(n)/DP(w) higher than 5.5/6.0 contained reducing groups. Trehalose contained no reducing groups. Freeze drying of an alkaline phosphatase solution without protectant induced an almost complete loss of the activity of the protein. In contrast, when inulins with a DP(n)/DP(w) higher than 5.5/6.0 or trehalose were used as stabilizer, the activity was fully maintained, also after subsequent storage for 4 weeks at 20 degrees C and 0, 45, or 60% RH, respectively. The stabilizing capacities of inulin with a lower DP and glucose were substantially less pronounced. After storage at 60 degrees C for 6 days, the activity of freeze dried samples containing inulins with a DP(n)/DP(w) higher than 5.5/6.0 was still about 50% whereas the activity of samples containing inulin with a lower DP, glucose, or trehalose was completely lost. It is concluded that inulins with a DP(n)/DP(w) higher than 5.5/6.0 meet the physico-chemical characteristics to successfully act as protectants for proteins. The stabilizing potential of these inulins was clearly shown using alkaline phosphatase as a model protein.
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