Characterizing the Freeze-Drying Behavior of Model Protein Formulations
Characterizing the Freeze-Drying Behavior of Model Protein Formulations
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DOI：
10.1208/s12249-010-9530-9
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发表时间：
2010-12-01
影响因子：
3.3
通讯作者：
Sinha, Sandipan
中科院分区：
文献类型：
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作者：
Lewis, Lavinia M.;Johnson, Robert E.;Sinha, Sandipan
三种模型蛋白质，即溶菌酶、BSA和IgG的冷冻干燥行为已经在两种不同的初级干燥条件（搁板温度分别为-25 ° C和+25 ° C）下在无定形制剂中使用多种技术进行了研究。使用测压温度测量来表征初级干燥期间的产品温度（T（pr））、升华速率和产品电阻（R（p））。生物物理技术，如圆二色谱，荧光，傅里叶变换红外光谱被用来研究蛋白质构象。使用尺寸排阻色谱法监测高分子量物质（HMWS）随储存时间的形成，并使用扫描电子显微镜研究滤饼形态。三种蛋白质的冻干行为差异在较高蛋白质浓度下更明显，其中蛋白质显著影响制剂基质的行为。然而，这些差异在积极模式下最小化，并且在辅料主导冻干行为的较低蛋白浓度下不显著。在两种干燥条件之间以及所研究的三种蛋白质之间观察到饼状物形态的差异。然而，使用两种不同的一级干燥条件生成的蛋白质饼的稳定性和蛋白质结构是等同的。
The freeze-drying behavior of three model proteins, namely, lysozyme, BSA, and IgG, has been studied using a variety of techniques under two different primary drying conditions (shelf temperatures of -25 degrees C and +25 degrees C, respectively) in an amorphous formulation. Manometric temperature measurements were used to characterize product temperature (T(pr)), sublimation rates, and product resistance (R(p)) during primary drying. Biophysical techniques such as circular dichroism, fluorescence, and Fourier transform infrared spectroscopy were used to study protein conformation. Size exclusion chromatography was used to monitor the formation of high-molecular-weight species (HMWS) over time on storage, and cake morphology was studied using scanning electron microscopy. The differences in the freeze-drying behavior of the three proteins were more evident at higher protein concentrations, where the protein significantly influences the behavior of the formulation matrix. However, these differences were minimized in the aggressive mode and were insignificant at lower protein concentrations where excipients dominated the freeze-drying behavior. Differences in cake morphology were observed between the two drying conditions employed as well as between the three proteins studied. The stability and the protein structure, however, were equivalent for the protein cakes generated using the two different primary drying conditions.