Evidence of Partial Unfolding of Proteins at the Ice/Freeze-Concentrate Interface by Infrared Microscopy
Evidence of Partial Unfolding of Proteins at the Ice/Freeze-Concentrate Interface by Infrared Microscopy
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DOI：
10.1002/jps.21843
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发表时间：
2009-09-01
影响因子：
3.8
通讯作者：
Nail, Steven L.
中科院分区：
文献类型：
--
作者：
Schwegman, J. Jeff;Carpenter, John F.;Nail, Steven L.
本研究的目的是使用红外光谱与冷冻干燥阶段相结合，以更好地了解由于与冷冻相关的应力而导致蛋白质完整性丧失的机制。在部分冷冻的系统中，一式三份地收集冰晶之间的间隙空间和通过冰晶的红外光谱。在存在和不存在添加的表面活性剂（聚山梨酯80）的情况下，收集乳酸脱氢酶（LDH）和人免疫球蛋白（IgG）的光谱。在间隙空间中收集的光谱，远离冰晶的表面，是非常相似的光谱从初始溶液中收集的表面活性剂的存在下，无论。通过冰晶收集的光谱，没有添加表面活性剂，显着不同的光谱从初始溶液中收集。在这些光谱中存在分子间P-片层结构（聚集体的主要成分）的谱带特征的增加。两种蛋白质制剂中的表面活性剂的存在下，导致在冰晶表面上的蛋白质的光谱中的分子间P-片层信号的减少。此外，当在冷冻之前将表面活性剂添加到制剂中时，最初在冰晶表面上损失的LDH的大部分天然状态结构恢复。(c)2009 Wiley-Liss，Inc.和美国药学协会药物科学杂志98：3239-3246，2009
The goal of this research was to use infrared spectroscopy in combination with a freeze drying stage to gain a better understanding of the mechanism of loss of protein integrity due to the stresses associated with freezing. Infrared spectra were collected in triplicate for the interstitial space between ice crystals and through ice crystals in a partially frozen system. Spectra were collected for lactate dehydrogenase (LDH) and human immune globulin (IgG) both in the presence and absence of an added surfactant (polysorbate 80). Spectra collected in the interstitial space, distant from the surface of ice crystals, were very similar to spectra collected from the initial solution regardless of the presence of a surfactant. Spectra collected through ice crystals, without added surfactant, were significantly different than spectra collected from the initial solution. An increase in bands characteristic of intermolecular P-sheet structures (main component of aggregates) were present in these spectra. The presence of surfactant in both protein formulations resulted in a decrease in intermolecular P-sheet signals in spectra of the proteins on the ice crystal surface. Additionally, much of the native state structure of LDH initially lost on the surface of ice crystals returned when surfactant was added to the formulation prior to freezing. (c) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3239-3246, 2009