The Effect of Mannitol Crystallization in Mannitol-Sucrose Systems on LDH Stability During Freeze-Drying
The Effect of Mannitol Crystallization in Mannitol-Sucrose Systems on LDH Stability During Freeze-Drying
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DOI：
10.1002/jps.23173
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发表时间：
2012-07-01
影响因子：
3.8
通讯作者：
Gieseler, Henning
中科院分区：
文献类型：
--
作者：
Al-Hussein, Anas;Gieseler, Henning
本研究的目的是研究甘露醇结晶和退火对冷冻干燥过程中乳酸脱氢酶（LDH）稳定性的影响。为此，对甘露醇与蔗糖的重量比不同的蛋白质配方进行了有退火和不退火的冷冻干燥。根据差示扫描量热数据计算产品结晶度。通过测量模型蛋白冷冻干燥后LDH的活性恢复来评估蛋白质的稳定性，并通过分析蛋白质二级结构来评估蛋白质的结构稳定性。LDH在退火样品中的稳定性较低，产物结晶度与模型蛋白的稳定性之间存在相关性。在冷冻干燥过程中，甘露醇结晶对LDH的不稳定作用可能归因于甘露醇从含有蛋白质的无定形相中去除。此外，甘露醇晶体的形成似乎促进了蛋白质在冷冻干燥过程中的界面变性。在配方中加入0.01%的Tween 80，大大提高了LDH抗甘露醇结晶应力的结构和功能稳定性。(3)中国医药科学，2012,(2):1 - 4
The objective of this study was to investigate the influence of mannitol crystallization and the effect of annealing on the stability of lactate dehydrogenase (LDH) during freeze-drying. For this purpose, protein formulations with different weight ratios of mannitol to sucrose were freeze-dried with and without annealing. Product crystallinity was calculated based on differential scanning calorimetry data. Protein stability was evaluated both functionally by measuring the activity recovery of the model protein LDH after freeze-drying, and structurally by analyzing the protein secondary structure. LDH showed lower stability in annealed samples, and a correlation could be established between the extent of product crystallinity and the stability of the model protein. The destabilizing effect of mannitol crystallization on LDH during freeze-drying can likely be attributed to removal of mannitol from the amorphous phase containing the protein. In addition, the formation of mannitol crystals seemed to enhance the interfacial denaturation of protein during freeze-drying. The use of 0.01% of Tween 80 in the formulation greatly improved the structural and functional stability of LDH against stresses induced by mannitol crystallization. (C) 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 101: 2534-2544, 2012