Kizuki Shinji, Wang Zekun, Yamauchi Satoru, Torisu Tetsuo, Uchiyama Susumu
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Formulation Research Lab, Taiho Pharmaceutical Co., Ltd, 224-2 Ebisuno, Hiraishi, Kawauchi-Cho, Tokushim, 771-0194, Japan.
AAPS J. 2025 Jan 27;27(1):34. doi: 10.1208/s12248-025-01014-z.
Protein aggregates and particles in biopharmaceuticals can induce adverse immune responses in patients. Thus, suppression of the formation of protein aggregates and particles is important for the successful development of therapeutic proteins. Mechanical stresses, including agitation, are widely recognized as stress factors that generate protein aggregates and particles. However, although refrigerators and storage chambers generate weak vibration, there have been no studies of the impact of such weak vibration on aggregate and particle formation during storage. In this study, monomer loss and aggregate formation of a CTLA4-Ig were evaluated during storage in a refrigerator (having a vibration acceleration less than 0.006 G) with or without three vibration isolators. The vibration isolators reduced the vibration acceleration, thereby decreasing the rate of monomer loss and nanometer-sized aggregate formation. The increase in the aggregation rate due to the weak vibration was not mitigated by adding poloxamer 188 or eliminating the air-liquid interface, which are processes known to be effective in preventing protein aggregation due to mechanical stresses. Thus, reducing vibration should be an effective way to mitigate the risk of aggregate formation.
生物制药中的蛋白质聚集体和颗粒可在患者体内引发不良免疫反应。因此，抑制蛋白质聚集体和颗粒的形成对于治疗性蛋白质的成功开发至关重要。包括搅拌在内的机械应力被广泛认为是产生蛋白质聚集体和颗粒的应激因素。然而，尽管冰箱和储存室会产生微弱振动，但尚未有关于这种微弱振动对储存期间聚集体和颗粒形成影响的研究。在本研究中，评估了在有或没有三个隔振器的冰箱（振动加速度小于0.006 G）中储存期间CTLA4-Ig的单体损失和聚集体形成情况。隔振器降低了振动加速度，从而降低了单体损失率和纳米级聚集体形成率。添加泊洛沙姆188或消除气液界面并不能减轻由于微弱振动导致的聚集率增加，而这些方法已知对防止机械应力导致的蛋白质聚集有效。因此，减少振动应该是降低聚集体形成风险的有效方法。
