Department of Applied Science and Technology, Politecnico di Torino, 24 corso Duca degli Abruzzi, Torino 10129, Italy.
Standardisation Science Section, Analytical and Biological Sciences Division, National Institute for Biological Standards and Control, Blanche Lane, Medicines & Healthcare Products Agency, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK.
J Pharm Sci. 2020 Jan;109(1):254-263. doi: 10.1016/j.xphs.2019.04.014. Epub 2019 Apr 17.
In the present work, the effect of controlled nucleation on the stability of human growth hormone (hGH) during freeze-drying has been investigated. More specifically, the vacuum-induced surface freezing technique has been compared to conventional freezing, both with and without an annealing step. Size exclusion chromatography and cell-based potency assays have been used to characterize the formation of soluble aggregates and the biological activity of hGH, respectively. The results obtained indicate that controlled nucleation has a positive effect on both cycle performance and product homogeneity because of the formation of bigger ice crystals, and characterized by a narrower dimensional distribution. From the point of view of hGH stability, we observed that vacuum-induced surface freezing is not detrimental to the biological activity of the protein, or aggregate formation. In addition, the effect of 2 different formulations, including trehalose or cellobiose, on protein preservation was also considered for this study.
在本工作中，研究了控制成核对人生长激素（hGH）在冷冻干燥过程中稳定性的影响。更具体地说，比较了真空诱导表面冻结技术与传统冻结，包括有和没有退火步骤的情况。尺寸排阻色谱和基于细胞的效价测定分别用于表征可溶性聚集体的形成和 hGH 的生物活性。所得结果表明，由于形成更大的冰晶，并且具有更窄的尺寸分布，控制成核对循环性能和产品均一性具有积极影响。从 hGH 稳定性的角度来看，我们观察到真空诱导表面冻结不会损害蛋白质的生物活性或聚集体的形成。此外，还考虑了 2 种不同配方（海藻糖或纤维二糖）对蛋白质保存的影响。
