Surface adsorption of recombinant human interferon-γ in lyophilized and spray-lyophilized formulations
Surface adsorption of recombinant human interferon-γ in lyophilized and spray-lyophilized formulations
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DOI：
10.1002/jps.10135
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发表时间：
2002-06-01
影响因子：
3.8
通讯作者：
Randolph, TW
中科院分区：
文献类型：
--
作者：
Webb, SD;Golledge, SL;Randolph, TW
将重组人干扰素-γ (rhIFN-γ) 冻干或喷雾冻干于含 9.5% 海藻糖、0.12% 聚山梨酯 20 的 10 mM 磷酸钾溶液中，pH 7.5。我们测量了喷雾、冻融、干燥和重构后可溶性蛋白质的回收率。红外光谱显示所有干燥粉末中的 rhIFN-γ 二级结构均与天然类似。使用电子光谱进行化学分析、飞行时间二次离子质谱、X 射线衍射和气体吸附等温线对粉末进行表征。 rhIFN-γ在喷雾过程中吸附在空气/液体界面上，在冻干过程中吸附在冰/液体界面上。冰/液体界面处的rhIFN-γ浓度约为空气/液体界面处吸附浓度的四分之一。添加 0.12% 聚山梨醇酯 20 降低了两个界面处 rhIFN-γ 的浓度。飞行时间二次离子质谱检测到冻干粉末表面存在聚山梨醇酯 20。与喷雾冻干样品相比，冻干样品干燥速度更慢，但重构速度更快。 rhIFN-gamma在雾化后聚集，但在0.12%聚山梨酯20中聚集减少。添加0.12%聚山梨酯20减少了蛋白质表面吸附并减少但没有完全阻止聚集。暴露于冰/液界面后发生不显着的聚集，但随后的干燥和重构引起聚集。大部分聚集是由于喷雾冻干或冻干过程中形成的空气-液体和固体-空气界面处的吸附造成的。 (C) 2002 Wiley-Liss, Inc. 和美国制药协会 J Pharm Sci 91:1474-1487, 2002。
Recombinant human interferon-gamma (rhIFN-gamma) was lyophilized or spraylyophilized in 9.5% trehalose, 0.12% polysorbate 20 in 10 mM potassium phosphate, pH 7.5. We measured recovery of soluble protein after spraying, freeze-thawing, and drying and reconstitution. Infrared spectroscopy showed rhIFN-gamma secondary structure to be native-like in all dried powders. Powders were characterized using electron spectroscopy for chemical analysis, time-of-flight secondary ion mass spectroscopy, X-ray diffraction, and gas adsorption isotherms. rhIFN-gamma adsorbed at air/liquid interfaces during spraying, and to ice/liquid interfaces during lyophilization. The concentration of rhIFN-gamma at ice/liquid interfaces was approximately one-fourth that adsorbed at air/liquid interfaces. Addition of 0.12% polysorbate 20 reduced the concentration of rhIFN-gamma at both interfaces. Time-of-flight secondary ion mass spectroscopy detected polysorbate 20 on surfaces of lyophilized powders. Lyophilized samples dried more slowly but reconstituted more quickly than spray-lyophilized samples. rhIFN-gamma aggregated after nebulization, but aggregation decreased in 0.12% polysorbate 20. Addition of 0.12% polysorbate 20 reduced protein surface adsorption and decreased but did not completely prevent aggregation. Insignificant aggregation occurred after exposure to ice/liquid interfaces, but subsequent drying and reconstitution caused aggregation. The majority of the aggregation is due to adsorption at air-liquid and solid-air interfaces formed during spray-lyophilization or lyophilization. (C) 2002 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 91:1474-1487, 2002.