Subvisible Particles Derived by Dropping Stress Enhance Anti-PEG Antibody Production and Clearance of PEGylated Proteins in Mice
Subvisible Particles Derived by Dropping Stress Enhance Anti-PEG Antibody Production and Clearance of PEGylated Proteins in Mice
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降低压力产生的亚可见颗粒可增强小鼠体内抗 PEG 抗体的产生和聚乙二醇化蛋白的清除
DOI：
10.1016/j.xphs.2022.01.023
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发表时间：
2022
影响因子：
3.8
通讯作者：
Tsutsumi Yasuo
中科院分区：
文献类型：
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作者：
Nakajima Takaki;Nagano Kazuya;Fukuda Yuka;Ishima Yu;Shibata Hiroko;Isaka Ryo;Zhang Tian-qi;Haga Yuya;Higashisaka Kazuma;Tsujino Hirofumi;Ishida Tatsuhiro;Ishii-Watabe Akiko;Tsutsumi Yasuo
聚乙二醇的生物偶联对蛋白质药物的开发非常重要，因为它提高了生物稳定性。相比之下，包括聚乙二醇化的蛋白质在内的蛋白质容易受到物理化学压力的影响。特别是，溶液中的蛋白质药物如果在运输过程中暴露在下降的压力下，可能会形成聚集体或亚可见颗粒。然而，许多聚乙二醇化的研究都集中在聚乙二醇化的有用性上，例如在血液中的半衰期的延长，以及聚乙二醇化蛋白质在下降压力下的物理性质或生物反应的变化还没有被探索。在这里，我们制备了四个不同长度(5或20 kDa)和不同数目(大[L]或小[S])的聚乙二醇化卵清蛋白(PEGylated OVA)分子，分析了在滴加压力下亚可见颗粒的形成，并考察了它们对抗体产生和清除的影响。在滴加胁迫下，20 kDa聚乙二醇卵蛋白(S)和L卵蛋白溶液的聚集颗粒浓度约为卵蛋白溶液的3倍。此外，滴注20 kDa的聚乙二醇卵蛋白可诱导小鼠产生抗聚乙二醇单抗，清除聚乙二醇卵蛋白。作为一种机制，下降应激可促进巨噬细胞摄取20 kDa的PEGOVA(L)。这些发现可以为适当的运输条件提供见解，以确保聚乙二醇化蛋白药物的质量。
Bioconjugation with polyethylene glycol (PEG) is important for protein drug development as it has improved biological stability. In contrast, proteins including PEGylated ones are susceptible to physicochemical stresses. Particularly, protein drugs in solution may form aggregates or subvisible particles if they are exposed to dropping stress during transportation. However, many PEGylation studies have focused on its usefulness, such as the extension of half-life in blood, and changes in the physical properties or biological responses of PEGylated proteins under dropping stress remain unexplored. Here, we prepared four PEGylated ovalbumin (PEG-OVA) molecules conjugated with different lengths (5 or 20 kDa) and numbers (large [L] or small [S]) of PEG, analyzed the formation of subvisible particles under dropping stress, and examined their impact on antibody production and clearance. Under dropping stress, the aggregated particle concentration of 20 kDa PEG-OVA (S) and (L) solutions was approximately 3-fold that of the OVA solution. Moreover, administration of 20 kDa PEG-OVA with dropping stress induced anti-PEG antibody production and clearance of PEG-OVA. As a mechanism, dropping stress could enhance the uptake of 20 kDa PEG-OVA (L) by macrophages. These findings could provide insights into proper transportation conditions to ensure the quality of PEGylated protein drugs.