Inulin glasses for the stabilization of therapeutic proteins
Inulin glasses for the stabilization of therapeutic proteins
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DOI：
10.1016/s0378-5173(00)00677-3
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发表时间：
2001-03-14
影响因子：
5.8
通讯作者：
Frijlink, HW
中科院分区：
文献类型：
--
作者：
Hinrichs, WLJ;Prinsen, MG;Frijlink, HW
糖玻璃广泛用于在干燥和随后的储存过程中稳定蛋白质。成功充当保护者。糖应具有高玻璃化转变温度（Tg）。吸湿性差，结晶速度低，不含还原基团。当设想冷冻干燥作为干燥方法时，优选冷冻浓缩级分的相对高的Tg(Tg')。在本研究中，调查了菊粉是否满足这些要求。对不同聚合度（DP）的菊粉进行了评估。海藻糖玻璃用作阳性对照。发现数/重平均DP(DPn/DPw)高于5.5/6.0的菊粉的Tg和Tg'高于海藻糖玻璃的Tg和Tg'。此外，菊粉玻璃表现出与海藻糖玻璃相似的吸湿性，但结晶速度较慢。 DPn/DPw 高于 5.5/6.0 的菊粉中，只有不到 6% 的糖单元含有还原基团。海藻糖不含还原基团。不含保护剂的碱性磷酸酶溶液冷冻干燥导致蛋白质活性几乎完全丧失。相比之下，当使用DPn/DPw高于5.5/6.0的菊粉或海藻糖作为稳定剂时，在随后分别在20℃和0、45或60%RH下储存4周后，活性也得到完全维持。 DP 较低的菊粉和葡萄糖的稳定能力明显不那么明显。 60℃保存6天后，DPn/DPw高于5.5/6.0的菊粉冻干样品的活性仍约为50%，而DP较低的菊粉、葡萄糖或海藻糖的样品活性完全丧失。结论是，DPn/DPw高于5.5/6.0的菊粉符合成功充当蛋白质保护剂的理化特性。使用碱性磷酸酶作为模型蛋白清楚地显示了这些菊粉的稳定潜力。 (C) 2001 Elsevier Science B.V. 保留所有权利。
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 (DPn/DPw) 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 DPn/DPw 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 DPn/DPw 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 degreesC 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 degreesC for 6 days, the activity of freeze dried samples containing inulins with a DPn/DPw 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 DPn/DPw higher than 5.5/6.0 meet the physicochemical characteristics to successfully act as protectants for proteins. The stabilizing potential of these inulins was clearly shown using alkaline phosphatase as a model protein. (C) 2001 Elsevier Science B.V. All rights reserved.